bioinformant worldwide, l.l.c....biomedical market research (e.g. market analysis for the stem cell...
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BIOINFORMANT WORLDWIDE, L.L.C. JUNE 2013
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Statements regarding potential markets for products and services
Anticipated drivers of future market growth
Assessment of competitors and potential competitors
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Assessment of outcomes and financial impacts
Aspects of the stem cell industry and related businesses
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BIOINFORMANT WORLDWIDE, L.L.C. JUNE 2013
SUMMARY OF METHODOLOGY
One characteristic that sets BioInformant apart is that our leadership team comes from a
"Bioinformatics" background – the science of collecting and analyzing complex genetic codes.
The field of bioinformatics involves developing precise methods and advanced tools for
understanding very large quantities of biological data, such as the complete set of genes within a
cell (genomics), the complete set of proteins in a cell (proteomics), and the complete set of
compounds or substrates that are formed through enzymatic reactions within living systems
(metabolomics).
All of these areas of data involve huge, complex data-sets that must be examined in order to
identify the critical information impacting the system at large. We're trained in applying these
advanced research techniques to the field of market research. Our skill set makes us uniquely
positioned to: 1) Define scope, definitions, and boundaries of the cord blood industry; 2) Collect
relevant data from within this expansive system; 3) Extract relevant and critical information; and
4) Identify key trends for purposes of predicting future activity within the industry.
Additionally, we are uniquely suited to managing complex search queries within large global
databases. Within the stem cell industry, this allows us to summarize critical trend rate
information from global databases such as scientific publication databases, clinical trial
databases, patent databases, research grant funding databases, transplant registries, and more.
In summary, bioinformatics is an interdisciplinary field that develops methods and tools for
processing complex biological data. Biomedical market research (e.g. market analysis for the
stem cell industry) is an interdisciplinary field that develops methods and tools for analyzing
complex industry data. When performed effectively, the fields share many, if not all, of the same
techniques.
Due to our expertise in the field of bioinformatics, BioInformant employs advanced techniques for
deriving stem cell market research. The following constitute the basis for our Research &
Analysis:
Preliminary Research: Examination of studies that need further confirmation by the scientific community, using extensive secondary research.
Fill-gap Research: Selectively sampled and focused primary research as a fill-gap strategy.
Historic Analysis - Primary Product(s): Comprehensive analysis of all data for each primary product market.
Historic Analysis - End-User Market(s): Historic analysis of all end-user industries/markets, requiring technology and market evaluations, growth projections, and market size estimation of end-user markets.
Historic Supply Chain/Raw Materials Analysis: Comprehensive analysis of data for each primary market segment.
Data Consolidation: Merging of historic end-user market data to yield consolidated primary market data.
Cross Linking: Comparison of primary market data (historic) with resulting end-user consolidated market data and calculation of the variance in percentages between data sets by year.
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Variance Determination: Placement of a median figure for each year with a tolerance range equal to twice the variance percentage. The resulting numbers are recorded.
Projections: Forward projection of end-user markets based upon historic growth, technology, market trends, and primary research from the market place.
Variance Factorization: Consolidation of projected end-user market data to yield derived primary market data. The data is adjusted to the historic variance determinations, as above. The resulting data is further verified by confirmatory primary research.
Confirmatory Primary Research: Presentation of resulting data from companies or individuals participating as research partners. Variations from derived data are adjusted to reflect primary research-based consensus.
Electronically Based End-User Surveys: Distribution and utilization of electronically based end-user surveys. Surveys are distributed to a comprehensive panel of individuals within market segment(s) of interest. Statistical analysis is performed on the user-response data.
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TABLE OF CONTENTS I. Abstract……………………………………………………………………………………………………………p.8 II. Overview…………………………………………………………………………………………………………p.10
A. Definition B. Alternative Nomenclature C. Criteria for Identifying
1. Physical Characteristics 2. Functional Attributes
D. Benefits Relative to Other Stem Cells
III. Isolation Sources………………………………………………………………………………………………p.16 A. Adult B. Fetal IV. Cell Types Derived from MSCs…………………………………………………………………………p.20
V. Applications…………………………………………………………………………………………………….p.21
A. Basic Research Applications 1. Directed Differentiation of MSCs into Intra-Mesenchymal Lineages 2. Directed Differentiation of MSCs into Extra-Mesenchymal Lineages
B. Supporting Hematopoietic Cells C. Gene Transfer Applications D. Cell Transplantation for Site-Specific Repair
1. Overview 2. Major Applications
a. Osteogenic Repair b. Myogenic and Myocardial Repair c. Neural Repair d. Pancreatic Repair
E. Topical Therapy and Use of MSCs in Wound Healing F. Use of MSC in 3-D Scaffolds for Tissue Engineering Applications G. Drug Delivery Applications: Engineered MSCs as specific carriers of anti-cancer
drugs H. Drug Screening I. MSC-mediated Inhibition of Immune Effector Cells
VI. Application Priorities………………………………………………………………………………………p.39 A. Overall B. By Segment 1. Academic 2. Biotechnology 3. Pharmaceutical
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VII. Companies Offering Human MSC Research Products………………………………………p.46 A. Abcam B. BD Biosciences C. Celprogen D. Cyagen Biosciences (China only) E. Life Technologies F. LGC Standards/ATCC G. Lonza H. Millipore I. Miltenyi Biotec J. PAA - The Cell Culture Company K. PromoCell L. SA Biosciences M. ScienCell Research Laboratories N. STEMCELLTechnologies O. Thermo Scientific P. R&D Systems
VIII. Companies Offering Rodent MSC Research Products………………………………………p.61
(Note: Eight companies offer rat MSC products: Cell Applications, Celprogen, Cyagen Biosciences, Genlantis, Millipore, Miltenyi Biotec, ScienCell Research Laboratories, and Trevigen. However, five were profiled in the previous section, leaving three to profile below as limited providers of rat MSC products.)
A. Cell Applications B. Genlantis C. Trevigen
IX. Analysis of MSC Research Activity……………………………………………………………………p.64
A. By Tissue Source of Origin B. By Clinical Application C. By Species Source D. By Geographical Region
X. Market Trend Analysis…………………………………………………………………………………….p.72
A. Scientific Publication Rate Analysis 1. Historical 10-Year Analysis 2. Future Five-Year Projection
B. Grant Trend Rate Analysis 1. Historical 10-Year Analysis 2. Future Five-Year Projection
C. Patent Analysis
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XI. Mesenchymal Stem Cells as Tools for Drug Discovery………………………………………p.85 A. Companies Developing Toxicology Screening Platforms for Stem Cells B. Potential Drug Screening Advantages of Mouse MSCs 1. Acquisition, Handling, and Differentiation Advantages
2. Genetic Manipulation of MSCs 3. Potential for Genetically Engineered MSC Reporter Lines XII. Serum Free Supplements for Use in MSC Research………………………………………….p.90
A. Invitrogen, Inc. B. Celprogen C. Millipore, Inc. D. Stem Cell Technologies E. US Patent 7109032: Serum-free medium for mesenchymal stem cells (Genoa, IT)
XIII. End-User Preferences (Scientist Panel)……………………………………………………………p.95
A. Source Preference B. Species Preference C. MSC Product Preferences D. Most Commonly Used MSC Antibodies E. Common Terms Used for Online Product Search
XIV. Research Product Opportunities and Suggestions…………………………………………p.104
A. Rat MSCs Derived from Synovium B. Novel MSC Differentiation Kits C. Mesenchymal Stem Cell Expansion Media D. Antibodies to Mesenchymal Stem Cell Markers E. Mesenchymal Growth Factors F. Monoclonal Antibodies and Kits for Phenotyping MSCs G. Chips for MicroArray Analysis of Gene Expression in MSCs
XV. Potential Labs/Customers……………………………………………………………………………p.114
A. General MSC Research Labs B. Osteogenic-Focused Research Labs C. Myogenic- and Myocardial-Focused Research Labs D. Adipogenic-Focused Research Labs E. Neuronal-Focused Research Labs F. Immunology-Focused Research Labs G. National Institutes of Health (NIH) Labs H. Commercial Labs
XVI. Conclusions………………………………………………………………………………………………….p.123
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I. ABSTRACT
Trend analysis of global grant activity, scientific publication rates, and patent
applications trends reveal that research activity involving mesenchymal stem cells
(MSCs) increased 112% from 2009 to 2010, and 116% from 2010 to 2011. Of most
interest is that this rate of growth accelerated throughout 2011, making mesenchymal
stem cells the fastest growing area of stem cell research.
MSCs are of therapeutic interest because they represent a population of cells that have
the potential to treat a wide range of acute and degenerative diseases. MSCs are also
advantageous over other stem cells types for a variety of reasons: they avoid the ethical
issues that surround embryonic stem cell research, and repeated studies have found
MSCs to be immuno-privileged, which make them an advantageous cell type for
allogenic transplantation. MSCs reduce both the risks of rejection and complications of
transplantation. Recently, there have been advances in the use of autologous
mesenchymal stem cells to regenerate human tissues, including cartilage, meniscus,
tendons, bone fractures, and more.
Because mesenchymal stem cell researchers represent a rapidly growing, well-funded
research community, this report presents strategies for research supply companies to
use to develop product lines that will be of high value to this community.
It is also important for bio-pharmaceutical and pharma companies interested in
mesenchymal stem cell therapy applications to understand underlying market forces,
and in particular, to consider progressive areas of MSC research as opportunistic areas
for drug and therapy development. This report presents a range of topics of interest to
these companies as well, including how advances in MSC research can reveal potential
new drug targets, improve methods of drug delivery, and provide personalized
treatment strategies.
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Highlights of this global strategic report include:
Advances in MSC research applications
Research priorities by market segment
Detailed characterization of the market, with trend assessment
Historical and future five-year trend projections
Exploration of recent product evolution and projected directions for
development
Data on end-user interests, needs and technical preferences (Scientist Panel)
Individual labs and end-users of MSC research products
Exploration of profit opportunities
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II. OVERVIEW
A. Definition
Broadly defined, mesenchymal stem cells (MSCs) are multi-potent stem cells that
can differentiate into a diverse range of cell types.
Specifically defined, mesenchymal stem cells are non-hematopoietic stromal cells
that are capable of differentiating into, and assisting in the repair of, tissues of both
intra-mesenchymal and extra-mesenchymal lineages. Mesenchyme is a form of
loose connective tissue within an embryo that contains undifferentiated cells
capable of differentiating into bone, cartilage, connective tissue, and cells of the
lymphatic and circulatory systems of an adult being. As such, examples of intra-
mesenchymal lineages include osteoblasts, chondrocytes, myocytes, and specialized
cells of the circulatory and lymphatic systems. Non-mesenchymal lineages include
beta-pancreatic islet cells, neural cells, and cells not associated with the circulatory,
lymphatic, or musculo-skeletal systems.
While not immortal, MSCs have the ability to expand significantly in a culture while
retaining their growth and multi-lineage potential. MSCs are identified by the
expression of many molecules, including CD105 (SH2) and CD73 (SH3/4), and are
negative for the hematopoietic markers CD34, CD45, and CD14.1
B. Alternative Nomenclature
Despite the definitions above, there is still some controversy over what constitutes a
“true” mesenchymal stem cell. Debate also exists as to what is the best and most
accurate terminology to be used for naming purposes.
1 Chamberlain G, Fox J, Ashton B, Middleton J. Concise review: Mesenchymal stem cells: Their phenotype, differentiation capacity, immunological features, and potential for homing. Stem Cell 2007; 25: 2739-2749.
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This has resulted in the existence of alternative nomenclature for the cell type,
which include:
1. Mesenchymal Stem Cell
2. Marrow Stromal Cell
3. Multi-potent Stromal Cell
4. Colony-Forming Unit-fibroblasts (CFU-f)
To date, the terms Mesenchymal Stem Cell and Marrow Stromal Cell have been
used interchangeably within the research community. In truth, however, neither
term provides an ideal description.
Mesenchymal Stem Cell: As described above, mesenchyme is a form of loose
connective tissue within an embryo that contains undifferentiated cells capable of
differentiating into bone, cartilage, connective tissue, and cells of the lymphatic
system and circulatory systems. It is an embryonic connective tissue derived from
the mesoderm, one of the three primary germ layers. The purpose of mesenchyme
tissue is to differentiate into hematopoietic cells (forming the blood system), as well
as to form cells of the lymphatic and musculo-skeletal systems. However, because
MSCs do not differentiate into hematopoietic cells, this term is not suitably
descriptive.
Marrow Stromal Cell: On the other hand, stromal cells are connective tissue cells
that form the supportive structure for the functional cells of a tissue or organ. While
this is an accurate description for one function of MSCs, the term fails to encompass
several other qualities of MSCs, including their ability to differentiate into non-
mesenchymal lineages, their active role in tissue repair, and their ability to exert
control over the immune response.
Multi-potent Stromal Cell: Because MSCs can differentiate into a range of different
cell types, but do not have the ability to form an entire organ, some researchers
have also proposed use of the term Multi-potent Stromal Cell. While this term is
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compatible from a terminology perspective, it has not been met with the same
general use or appeal as the term “Mesenchymal Stem Cell.”
Colony-Forming Unit-fibroblasts: Furthermore, historically-based nomenclature
suggests use of another name. In 1924, morphologist Alex Maximow used
histological observations to identify a type of precursor cell within mesenchyme that
could develop into different types of blood cells;2 in the 1960s, scientists McCulloch
and Till discovered the clonal nature of those cells.3,4 An assay system
demonstrating clonogenic potential of multi-potent stromal cells was presented in
1974 by Friedenstein and colleagues, and in this system, these cells were referred to
as Colony-Forming Unit-fibroblasts (CFU-f).5
However, while there remains occasional use of alternative names, it is now
generally accepted within the scientific community that the term “mesenchymal
stem cell” is the proper term for use in publication involving peer review. In
addition, industry forces (including research supply companies and drug therapy
companies) have further propagated use of the term “mesenchymal stem cell” over
the past few years.
Thus, for purposes of marketing a product line to this research community, it is
recommended that the term “mesenchymal stem cell” be incorporated into the
product title. However, for purposes of optimal website traffic flow, pages that
feature MSC products should be designed to collect traffic flow resulting from
searches for all four of the descriptive names. A credible website designer will know
how to use “Keyword” Meta tags, image description and “Alt” data, and keyword
frequency, among other Search Engine Optimization (SEO) techniques, to optimize
product pages to collect this traffic. In addition, the abbreviations “MSC” and
“MSCs” should be considered for search optimization as well.
2 Sell S. Stem cell handbook 2003; 143. 3 Becker AJ, McCulloch EA, Till JE. Cytological demonstration of the clonal nature of spleen colonies derived from transplanted mouse marrow cells. Nature 1963; 197: 452–454. 4 Siminovitch L, McCulloch EA, Till JE. The distribution of colony-forming cells among spleen colonies. Journal of Cellular and Comparative Physiology; 1963; 62: 327–336. 5 Friedenstein AJ, Deriglasova UF, Kulagina NN, Panasuk AF, Rudakowa SF, Luria EA, Ruadkow IA. Precursors for fibroblasts in different populations of hematopoietic cells as detected by the in vitro colony assay method. Exp Hematol 1974; 2: 83–92.
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C. Criteria for Identifying
The criteria used to identify mesenchymal stem cells can also vary. Identification of
MSCs can be accomplished either through physical or functional attribute
assessment.
1. Physical Assessment
The first, and usually most convenient, approach is physical assessment.
Morphologically, MSCs have a small cell body with long, thin cell processes that
can give the cells a “stretched” appearance. The cell body contains a large,
round nucleus with a prominent nucleolus. The cells present with symmetrical
morphology.
MSCs tend to be broadly dispersed within an extracellular matrix (ECM) that
contains a few reticular fibrils but lacks other types of collagen fibrils.
2. Functional Attributes
A second approach for identifying mesenchymal stem cells is through functional
assessment. Criteria for identifying MSCs are presented below:
Able to be isolated by plastic adherence
Have multi-potential and multi-lineage capability (plasticity)
Distinctive Surface Markers, specifically: CD73, CD90, CD1056
Lack of Lineage-Specific Markers, including: CD34, CD14, CD457
Immuno-favorable characteristics that include little to no expression of
MHC class I antigens and very limited expression of MHC class II antigens8
6 Klingemann H, et al. Mesenchymal Stem Cells, Sources and Applications. Transfusion Medicine and Hemotherapy 2008; 35: 272-277. 7 Friedman R, Betancur M, Tuncer H, Boissel L, Klingemann H. Umbilical cord mesenchymal stem cells: Adjuvants for human cell transplantation. Biol Blood Marrow Transplant 2007; 13: 1477–1486. 8 Friedman R, Betancur M, Tuncer H, Boissel L, Klingemann H. Umbilical cord mesenchymal stem cells: Adjuvants for human cell transplantation. Biol Blood Marrow Transplant 2007; 13: 1477–1486.
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Lack of co-stimulatory molecules of the B7 family that are required to
initiate an immune response
In addition, functional assessment can include analyzing cells for characteristic
cytokine production or gene expression profiles. However, the difficulty with
this approach is that the “characteristic” cytokines produced or genes expressed
by MSCs can vary by source of origin. For instance, bone marrow-derived MSCs
and adipose-derived MSCs exhibit different, source-specific, cytokine
production.9
D. Benefits Relative to Other Stem Cells
Interestingly, MSCs have a range of benefits compared to other stem cell types, as
presented below:
1. Well-Characterized: MSCs are a well-characterized population of adult stem
cells, regarding which 20,673 scientific articles have been published.10
2. Non-Controversial: MSCs avoid the ethical issues of embryonic stem cells, as
they can be derived from sources that include adult bone marrow and
adipose tissue.
3. Diverse Differentiation Potential: MSCs can form a variety of cell types in the
laboratory, including those of both intra- and extra-mesenchymal lineage.
These cell types include: fat (adipocytes), bone (osteoblasts), skin (dermal
cells), nerve (neural cells), cartilage (chondrocytes), muscle (skeletal
myocytes), tendons (tenocytes), marrow stroma, ligaments, and more.
4. Ease of Growth in Culture: Advanced knowledge exists for how to grow
MSCs in culture, including protocols for isolation, expansion, and
differentiation.
9 Gonzalez-Rey1 E, et al. Human adipose-derived mesenchymal stem cells reduce inflammatory and T cell responses and induce regulatory T cells in vitro in rheumatoid arthritis. Ann Rheum Dis 2010; 69: 241-248. 10 Figure calculated using search of PubMed Database (http://www.ncbi.nlm.nih.gov/pubmed) for the search terms: ["Mesenchymal Stem Cell" OR "Mesenchymal Stem Cells"] OR ["Marrow Stromal Cell" OR "Marrow Stromal Cells"] OR ["Multi-potent Stromal Cell" OR "Multi-potent Stromal Cells"] OR ["Colony-Forming Unit-fibroblast" OR "Colony-Forming Unit-fibroblasts"]. Search encompassed four technical terms for the cell type, as well as variations in singular versus plural usage of terminology. Executed May 12, 2013.
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5. Flexible Propagation: MSCs can be grown and propagated in culture for
extended periods, without losing differentiation potential.
6. Clinically Relevant Volumes: Unlike many other types of adult stem cells,
MSCs can be acquired in the quantities required for clinical applications, as
knowledge exists for how to culture the cell type in 3D bioreactors. It is
understood that reduced oxygen conditions, along with available nutrients,
assist MSC expansion under bioreactor conditions.11
7. Role as Regulatory Cells: MSCs synthesize and secrete a variety of
macromolecules that are known regulators of hematopoietic and bone-
resorbing cells.12
8. Delivery of Gene Products: MSCs can take up exogenous DNA and keep
introduced genes, an attribute that may allow use of the cells in therapeutic
delivery of molecules to target regions of the body.
9. Favorable Immune Status: MSCs lack the co-stimulatory molecules of the B7
family that are required to initiate an immune response.13 This allows the
administration of MSC preparations across MHC barriers without concern for
immunological rejection or the need for immunosuppression, making MSCs a
universal stem cells source.
10. Commercially Available Research Tools: Currently, sixteen research supply
companies offer human MSC products and eight offer mouse MSC products,
making research tools for this cell type easily accessible.
11 Godara P, et al. Mini-review: Design of bioreactors for mesenchymal stem cell tissue engineering. J Chem Technol Biotechnol 2008; 83: 408–420. 12 Haynesworth S, Reuben D, Caplan A. Cell-based tissue engineering therapies: The influence of whole body physiology. Advanced Drug Delivery Reviews 1998; 33(1-2): 3-14. 13 Tipnis S, Viswanathan C, Majumdar A. Immunosuppressive properties of human umbilical cord-derived mesenchymal stem cells: Role of B7-H1 and IDO. Immunology and Cell Biology 2010; 88: 795-806.
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III. ISOLATION SOURCES
Mesenchymal stem cells can be derived from a diverse range of tissues. Originally,
Friedenstein et al. isolated MSCs from the bone marrow (BM) and stroma of the spleen
and thymus.14,15 MSCs have since been derived from tissues as varied as the brain,
spleen, liver, kidney, lung, bone marrow, muscle, thymus, and pancreas.16 However,
bone marrow aspirates are still considered to be the most convenient and enriched
source of MSCs.17
Fetal tissue also contains MSCs, with common sources including umbilical cord blood
and fetal placenta. These sources may represent ontogenetically younger MSCs.
Evidence exists that MSCs from fetal sources can undergo more cell divisions before
they reach senescence than MSCs from adult tissue.18
Variations at the genetic level have also been well documented for MSCs from different
sources,19 as have differences in the types of chemokines and cytokines the cells
produce.20
The most common adult sources of MSCs include:
Stroma of Bone Marrow: This was the first isolated source and remains the most
common one.
Adipose Tissue: Represents a readily available source of MSCs.
Dental Pulp: Interestingly, an extremely rich source for dental pulp MSCs is the
developing tooth bud of the mandibular third molar. MSCs derived from this
14 Friedenstein AJ, Gorskaja JF, Kulagina NN. Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 1976; 4(5): 267—274. 15 Friedenstein AJ, Piatetzky-Shapiro II, Petrakova KV. Osteogenesis in transplants of bone marrow cells. J Embryol Exp Morphol 1966; 16(3): 381—390. 16 da Silva Meirelles L, Chagastelles PC, Nardi NB. Mesenchymal stem cells reside in virtually all post-natal organs and tissue. Cell Science 2006. 17 Tuli R, Seghatoleslami MR, Tuli S, et al. A simple, high-yield method for obtaining multi-potential mesenchymal progenitor cells from trabecular bone. Mol Biotechnol 2003; 23(1): 37—49. 18 Klingemann H, et al. Mesenchymal Stem Cells – Sources and Clinical Applications. Transfus Med Hemother 2008; 35: 272-277. 19 Wagner W, Wein F, Seckinger A, et al. Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood. Exp Hematol 2005; 33: 1402–16. 20 Friedman R, Betancur M, Tuncer H, Boissel L, Klingemann H. Umbilical cord mesenchymal stem cells: Adjuvants for human cell transplantation. Biol Blood Marrow Transplant 2007; 13: 1477–1486.
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source demonstrate preferential capacity to differentiate into bone and
neurons.21,22
The most common fetal sources of MSCs include:
Umbilical Cord Blood: MSCs can be derived from umbilical cord vasculature,
although low count per volume usually means that expansion is required to
obtain practical quantities.
Wharton’s Jelly: MSCs are also found within this gelatinous substance within
the umbilical cord.
Placenta: MSCs can be derived from several placenta components, including
the chorion, amnion, and villous stroma.23
The table below presents adult tissue sources from which MSCs can be isolated. The
intriguing range of tissues encompasses everything from small vessels (such as
kidney glomeruli) to large blood vessels (including the aortic artery and vena cava).
In addition, it includes a large number of mesenchyme-derived tissue (bone
components, skeletal muscle, tendons, cartilage, and more), as well as non-
mesenchyme derived tissues (neural tissue).
21 Gronthos S, Brahim J, Li W, et al. Stem cell properties of human dental pulp stem cells. J Dent Res 2002; 8: 531–535. 22 Yu J, Wang Y, Deng Z, Li Y, Shi J, Jin Y. Odontogenic capability: bone marrow stromal stem cells versus dental pulp stem cells. Biol Cell 2007; 8: 465–474. 23 Portmann-Lanz CB. Placental mesenchymal stem cells as potential autologous graft for pre- and perinatal neuroregeneration. Am J Obstet Gynecol 2006; 194(3): 664-673.
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TABLE: Adult Sources for Mesenchymal Stem Cell (MSC) Isolation
24 De Bari C, Dell’Accio F, Vandenabeele F, et al. Skeletal muscle repair by adult human mesenchymal stem cells from synovial membrane. J Cell Biol 2003; 160(6): 909—918. 25 Alsalameh S, Amin R, Gemba T, Lotz M. Identification of mesenchymal progenitor cells in normal and osteoarthritic human articular cartilage. Arthritis Rheum 2004; 50(5): 1522—1532. 26 Friedenstein AJ, Gorskaja JF, Kulagina NN. Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 1976; 4(5): 267—274. 27 Friedenstein AJ, Piatetzky-Shapiro II, Petrakova KV. Osteogenesis in transplants of bone marrow cells. J Embryol Exp Morphol 1966; 16(3): 381—390. 28 Gronthos S, Brahim J, Li W, et al: Stem cell properties of human dental pulp stem cells. J Dent Res 2002; 8: 531–535. 29 Klingemann H, Matzilevich D, Marchand J. Mesenchymal Stem Cells – Sources and Clinical Applications. Transfus Med Hemother 2008; 35: 272-277. 30 Klingemann H, Matzilevich D, Marchand J. Mesenchymal Stem Cells – Sources and Clinical Applications. Transfus Med Hemother 2008; 35: 272-277. 31 Friedenstein AJ, Gorskaja JF, Kulagina NN. Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 1976; 4(5): 267—274. 32 Klingemann H, Matzilevich D, Marchand J. Mesenchymal Stem Cells – Sources and Clinical Applications. Transfus Med Hemother 2008; 35: 272-277. 33 Klingemann H, Matzilevich D, Marchand J. Mesenchymal Stem Cells – Sources and Clinical Applications. Transfus Med Hemother 2008; 35: 272-277. 34 Cuevas P, Carceller F, Garcia-Gomez I, et al. Bone marrow stromal cell implantation for peripheral nerve repair. Neurol Res 2004; 26(2): 230—232. 35 Young HE, Steele TA, Bray RA, et al. Human reserve pluripotent mesenchymal stem cells are present in the connective tissues of skeletal muscle and dermis derived from fetal, adult, and geriatric donors. Anat Rec 2001; 264(1): 51—62. 36 Young HE, Steele TA, Bray RA, et al. Human reserve pluripotent mesenchymal stem cells are present in the connective tissues of skeletal muscle and dermis derived from fetal, adult, and geriatric donors. Anat Rec 2001; 264(1): 51—62. 37 Young RG, Butler DL, Weber W, et al. Use of mesenchymal stem cells in a collagen matrix for Achilles tendon repair. J Orthop Res 1998; 16(4): 406—413. 38 Pountos I, Giannoudis P. Biology of mesenchymal stem cells. Injury, Int J Care Injured 2005; 365: S8-S12. 39 Friedenstein AJ, Gorskaja JF, Kulagina NN. Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 1976; 4(5): 267—274. 40 Pountos I, Giannoudis P. Biology of mesenchymal stem cells. Injury, Int J Care Injured 2005; 365: S8-S12. 41 Friedenstein AJ, Gorskaja JF, Kulagina NN. Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 1976; 4(5): 267—274. 42 Jones EA, English A, Henshaw K, et al. Enumeration and phenotypic characterization of synovial fluid multi-potential mesenchymal progenitor cells in inflammatory and degenerative arthritis. Arthritis Rheum 2004; 50(3): 817—827. 43 Salingcarnboriboon R, Yoshitake H, Tsuji K, et al. Establishment of tendon-derived cell lines exhibiting pluripotent mesenchymal stem cell-like property. Exp Cell Res 2003; 15: 289—300. 44 Tuli R, Seghatoleslami MR, Tuli S, et al. A simple, high-yield method for obtaining multi-potential mesenchymal progenitor cells from trabecular bone. Mol Biotechnol 2003; 23(1): 37—49. 45 Klingemann H, Matzilevich D, Marchand J. Mesenchymal Stem Cells – Sources and Clinical Applications. Transfus Med Hemother 2008; 35: 272-277. 46 Klingemann H, Matzilevich D, Marchand J. Mesenchymal Stem Cells – Sources and Clinical Applications. Transfus Med Hemother 2008; 35: 272-277.
SOURCE ABBREVIATION REFERENCE COMMENTS
Adipose Tissue AT-MSC 24 MSCs derived from this tissue do not differentiate well into chondrocytes.
Aortic Artery
Articular Cartilage 25
Bone Marrow BM-MSC 26, 27 BM-MSCs can be conveniently derived by bone marrow aspirate.
Dental Pulp DP-MSC 28
MSCs derived from this source demonstrate preferential capacity to differentiate into bone and neurons.
Kidney glomeruli 29
Liver L-MSC 30
Lung L-MSC 31
Neural Tissue N-MSC 32
SOURCE ABBREVIATION REFERENCE COMMENTS
Pancreas PMSC 33
Periostium - 34
Skeletal Muscle M-MSC 35
Dermis - 36,37
Stroma of Spleen S-MSC 38,39
Friedenstein et al. isolated MSCs from the stroma of the spleen and thymus as early as 1976.
Stroma of Thymus T-MSC 40,41
(See comment above.)
Synovium and Synovial Fluid -
42
Tendons - 43
Trabecular Bone - 44
Vena Cava - 45
Xiphoid Cartilage - 46
19
Similarly, the table below presents fetal sources from which MSCs can be isolated.
TABLE: Fetal Sources for Mesenchymal Stem Cell (MSC) Isolation
SOURCE ABBREVIATION REFERENCE COMMENTS
Umbilical Cord Vasculature CB-MSC 47, 48
Most common fetal source of MSCs. Fetal cord blood can be collected at birth and publicly or privately stored as a future source of MSCs.
Placenta - 49
Amnion A-MSC 50
Interestingly, MSCs derived from this source do not exhibit the ability to differentiate into adipocytes.
Wharton's Jelly WJ-MSC 51
Wharton's Jelly is a gelatinous substance found within the umbilical cord.
Fetal Tissues (Pancreas, Spleen, Thymus) - 52
MSCs from fetal tissues have been successfully differentiated into cells of osteogenic, chondrogenic, and adipogenic lineages.
47 Friedman R, Betancur M, Tuncer H, Boissel L, Klingemann H. Umbilical cord mesenchymal stem cells: Adjuvants for human cell transplantation. Biol Blood Marrow Transplant 2007; 13: 1477–1486. 48 Friedman R, Betancur M, Tuncer H, Boissel L, Klingemann H. Umbilical cord mesenchymal stem cells: Adjuvants for human cell transplantation. Biol Blood Marrow Transplant 2007; 13: 1477–1486. 49 Friedman R, Betancur M, Tuncer H, Boissel L, Klingemann H. Umbilical cord mesenchymal stem cells: Adjuvants for human cell transplantation. Biol Blood Marrow Transplant 2007; 13: 1477–1486. 50 Pountos I, Giannoudis P. Biology of mesenchymal stem cells. Injury, Int J Care Injured 2005; 36: S8—S12. 51 Friedman R, Betancur M, Tuncer H, Boissel L, Klingemann H. Umbilical cord mesenchymal stem cells: Adjuvants for human cell transplantation. Biol Blood Marrow Transplant 2007; 13: 1477–1486. 52 Ying Huab, et al. Isolation and identification of mesenchymal stem cells from human fetal pancreas. J Lab Clin Med 2003; 141(5): 342-349.
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IV. CELL TYPES DERIVED FROM MSCs
In addition, mesenchymal stem cells can differentiate into a diverse range of tissues,
including the many cell types shown in the table below.
TABLE: Differentiation Capacity of Mesenchymal Stem Cells
SOURCE REFERENCE
Osteoblasts 53
Chondrocytes 54,55
Adipocytes 56
Cardiac Myocytes 57
Fibroblasts 58
Myofibroblasts 59
Pericytes 60
Skeletal Myocytes 61
Tenocytes 62
Retinal Cells 63
Neural Cells 64
Astrocytes 65
Hepatocytes 66
Hematopoietic Supporting Stroma 67
Pancreatic Cells 68
53 Jones EA, Kinsey SE, English A, et al. Isolation and characterization of bone marrow multi-potential mesenchymal progenitor cells. Arthritis Rheum 2002; 46: 3349—3360. 54 Jones EA, Kinsey SE, English A, et al. Isolation and characterization of bone marrow multi-potential mesenchymal progenitor cells. Arthritis Rheum 2002; 46: 3349—3360. 55 Yoo JU, Barthel TS, Nishimura K, et al. The chondrogenic potential of human bone-marrow-derived mesenchymal progenitor cells. J Bone Joint Surg Am 1998; 80(12): 1745—1757. 56 Jones EA, Kinsey SE, English A, et al. Isolation and characterization of bone marrow multi-potential mesenchymal progenitor cells. Arthritis Rheum 2002; 46: 3349—3360. 57 Orlic D, Kajstura J, Chimenti S, et al. Bone marrow cells regenerate infarcted myocardium. Nature 2001; 410(6829): 701—705. 58 Dicker A, Le Blanc K, Astrom G, et al. Functional studies of mesenchymal stem cells derived from adult human adipose tissue. Exp Cell Res 2005; 308: 283—290. 59 Buckwalter JA. Articular cartilage: injuries and potential for healing. J Orthop Sports Phys Ther 1998; 28(4): 192—202. 60 Direkze NC, Forbes SJ, Brittan M, et al. Multiple organ engraftment by bone-marrow-derived myofibroblasts and fibroblasts in bone-marrow-transplanted mice. Stem Cells 2003; 21(5): 514—520. 61 Cuevas P, Carceller F, Garcia-Gomez I, et al. Bone marrow stromal cell implantation for peripheral nerve repair. Neurol Res 2004; 26(2): 230—232. 62 Pittenger M, Vanguri P, Simonetti D, Young R. Adult mesenchymal stem cells: potential for muscle and tendon regeneration and use in gene therapy. J Musculoskelet Neuronal Interact 2002; 2(4): 309—20. 63 Tomita M, Adachi Y, Yamada H, et al. Bone marrow-derived stem cells can differentiate into retinal cells in injured rat retina.Stem Cells 2002; 20(4): 279—283. 64 Long X, Olszewski M, Huang W, Kletzel M. Neural cell differentiation in vitro from adult human bone marrow mesenchymal stem cells.” Stem Cells Dev 2005; 14(1): 65—9. 65 Schor AM, Canfield AE, Sutton AB, et al. Pericyte differentiation. Clin Orthop Relat Res 1995; 313: 81—91. 66 Lee KD, Kuo TK, Whang-Peng J, et al. In vitro hepatic differentiation of human mesenchymal stem cells. Hepatology 2004; 40(6): 1275—1284. 67 Minguell JJ, Erices A, Conget P. Mesenchymal stem cells. Exp Biol Med 2001; 226(6): 507—520. 68 Chen LB, Jiang XB, Yang L. Differentiation of rat marrow mesenchymal stem cells into pancreatic islet beta-cells. World J Gastroenterol 2004; 10(20): 3016—3020.
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V. APPLICATIONS
MSCs have utility in wide range of applications, spanning from the basic (exploratory
research) to the dramatic (regenerative medicine and tissue engineering).
A. Basic Research Applications
To advance scientific progress with any cell type, a basic level of knowledge needs to
be acquired and shared. This knowledge includes developing protocols for cell
isolation, growth, expansion, and differentiation. It requires that characteristics of
the cells be observed to determine how they are affected by environmental
variation, such as differences in handling or local cues.
A significant amount has been learned about mesenchymal stem cells, as 20,673
scientific articles have been published about the cell type to date.69 However, there
remain areas of mesenchymal stem cell activity that are unclear, particularly when
the cells are considered for clinical application. These areas include how to guide
MSCs in vivo, how MSCs interact as implanted cells with local signals, and how MSCs
create immuno-modulatory (immune suppressing) effects.
For this reason, basic research activity with MSCs is critical. It furthers the ability of
medical researchers to safely and practically investigate MSCs as therapeutic agents
and consider them for use in applied fields, such as regenerative medicine, tissue
engineering, and drug delivery.
1. Directed Differentiation of Mesenchymal Stem Cells into Intra-Mesenchymal
Lineages
As described earlier, mesenchymal stem cells are multi-potent stem cells that
can differentiate into a variety of cell lineages. Since the 1960s when scientists
69 Figure calculated using search of Pubmed Database (http://www.ncbi.nlm.nih.gov/pubmed) for the search terms: ["Mesenchymal Stem Cell" OR "Mesenchymal Stem Cells"] OR ["Marrow Stromal Cell" OR "Marrow Stromal Cells"] OR ["Multi-potent Stromal Cell" OR "Multi-potent Stromal Cells"] OR ["Colony-Forming Unit-fibroblast" OR "Colony-Forming Unit-fibroblasts"]. Search encompassed four technical terms for the cell type, as well as variations in singular versus plural usage of terminology. Executed May 12, 2013.
22
Ernest McCulloch and James Till revealed the clonal nature of marrow-derived
mesenchymal cells, it has been understood that MSCs are characterized by
plasticity and that their fate can be determined by environmental cues.70
It is now evident that culturing marrow stromal cells in the presence of
osteogenic stimuli, such as ascorbic acid, inorganic phosphate, and
dexamethasone, can promote differentiation into osteoblasts.71 Alternatively,
the addition of Transforming Growth Factor-beta (TGF-b) can induce
chondrogenic markers.72 Myocyte and adipocyte differentiation can be similarly
induced.
Directed differentiation of autologous mesenchymal stem cells into intra-
mesenchymal lineages is an application that involves identifying pharmacological
and molecular pathways that drive MSC differentiation toward mesenchymal
derivatives in vitro. Its goal is to assist with predicting molecular mechanisms
that will control MSC differentiation in vivo.
In particular, there is a need to develop reliable methods for directing the
differentiation of human mesenchymal stem cells (hMSC) during regenerative
medicine applications. Most research in this area is focused on embedding
mesenchymal cells into defined protein microenvironments and tracking
directed differentiation through cell morphology, gene expression, and cell
activity.
2. Directed Differentiation of MSCs into Extra-Mesenchymal Lineages
Directed differentiation of autologous mesenchymal stem cells into extra-
mesenchymal lineages is another interesting area of stem cell biology, with the
potential to repair tissues where resident stem cells are not accessible. The
70 Becker AJ, McCulloch EA, Till JE. Cytological demonstration of the clonal nature of spleen colonies derived from transplanted mouse marrow cells. Nature 1963; 197: 452–454. 71 Ye CP, et al. Culture media conditioned by heat-shocked osteoblasts enhances the osteogenesis of bone marrow-derived mesenchymal stromal cells. Cell Biochem Funct 2007; 25(3): 267-276. 72 Mehlhorn AT, et al. Mesenchymal stem cells maintain TGF-beta-mediated chondrogenic phenotype in alginate bead culture. Tissue Eng 2006 Jun; 12(6): 1393-1403.
23
potential to differentiate MSCs into neuronal cells is a possibility that has already
been demonstrated73 and an area that continues to be of significant clinical
interest.
It has also been demonstrated that mesenchymal stem cells can differentiate
into beta-pancreatic islet cells74. In 2004, Chen and colleagues explored the
possibility that bone marrow mesenchymal stem cells could differentiate in vitro
into functional islet-like cells. His research team discovered that when rat MSCs
were isolated and cultured, passaged MSCs could be induced to differentiate
into typical islet-like clustered cells. Insulin mRNA and protein expressions were
positive in populations of the differentiated cells, and nestin could be detected in
pre-differentiated cells. Furthermore, insulin excreted from differentiated MSCs
was much higher than that from pre-differentiated cells, and injecting the
differentiated MSCs into diabetic rats supported down-regulation of glucose
levels in test subjects. As such, transplantation of MSC-derived islet-like
functional cells may eventually be used in clinical applications for the treatment
of diabetes.
However, some scientists believe that experimentation in this area is
inconclusive and that substantial research needs to be done before human
studies are untaken that involve mesenchymal stem cell-derived neural- or beta-
pancreatic islet cells. While it is true that mechanisms for extra-mesenchymal
differentiation are not well understood, cautious optimism does seem
appropriate.
One reason that research in this area will continue is that there is significant
medical need for extra-mesenchymal lineage cell types, most notably neural cells
that could be used for the treatment of degenerative brain diseases and hepatic
cells that could be used for diabetic therapy applications. In addition, research in
this area will be driven by the wide range of benefits associated with MSCs,
73 Engler AJ, Sen S, Sweeney HL, Discher DE. Matrix Elasticity Directs Stem Cell Lineage Specification. Cell 2006; 126(4): 677-689. 74 Chen LB, Jiang XB, Yang L. Differentiation of rat marrow mesenchymal stem cells into pancreatic islet beta-cells. World J Gastroenterol 2004; 10(20): 3016–3020.
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which include ease of acquisition from a range of adult tissues, flexible methods
for growth in culture, and expansion capabilities that allow for clinically relevant
quantities to be obtained.
In 2012 (January 1, 2012 - January 1, 2013), rates of research exploring directed
differentiation of MSCs toward extra-mesenchymal lineages comprised only
7.9% of MSC-directed differentiation research, while rates of research exploring
directed differentiation of MSCs toward intra-mesenchymal lineages comprised
92.1 percent.75 (See graph below.)
Table: Breakdown of MSC Directed Differentiation Research (January 1, 2012 – January 1, 2013)
Type of Research Percent
Intra-Mesenchymal Lineage Research 7.9%
Extra-Mesenchymal Lineage Research 7.9%
TOTAL 100.0%
92.1%
7.9%
Breakdown of MSC Directed Differentiation Research (January 1, 2012 – January 1, 2013)
Intra-Mesenchymal LineageResearch
Extra-Mesenchymal LineageResearch
Additionally, there are interesting trends for Google searches for the phrase “Mesenchymal Stem Cell” or “Mesenchymal Stem Cells;” the results, in conjunction with a Deep Web76 search, are presented below.
75 Lineage-specific search phrase analysis of PubMed (http://www.ncbi.nlm.nih.gov/pubmed) and Highwire Press (http://highwire.stanford.edu) publication databases. Test Interval: Jan 1, 2012 – Jan 1, 2013. 76 The term "Deep Web" was coined by BrightPlanet, an Internet search technology company that specializes in searching Deep Web content. In their 2001 white paper, “The Deep Web: Surfacing Hidden Value,” BrightPlanet noted that the deep Web was growing more quickly than the surface Web and that the quality of the content within it is significantly higher than the vast majority of
25
YEAR # OF GOOGLE SEARCH RESULTS # OF SEARCH RESULTS (GOOGLE + DEEP WEB)
2003 8,980 12,681
2004 10,400 14,107
2005 13,600 18,405
2006 20,300 25,896
2007 34,800 39,891
2008 44,000 53,909
2009 57,300 68,122
2010 70,800 95,921
2011 104,000 141,648
2012 134,071 207,832
0
50,000
100,000
150,000
200,000
250,000
2003 2004 2005 2006 2007 2008 2009 2010 2011 2012
# of Google Search Results # of Search Results (Google + Deep Web)
surface Web content. It is estimated that 95% of the Deep Web can be accessed through specialized search. The Deep Web contains over 980 billion pages, while the Surface Web which is composed of only 35 billion pages. In particular, Deep Web search allows a user access to product/service pages; advertiser bids and posts; and databases including government, business and university databases. In this analysis, the information that was quantified was search volume for the search terms: “Mesenchymal Stem Cell" and "Mesenchymal Stem Cells."
26
B. Supporting Hematopoietic Cell Applications
Mesenchymal stem cells also play a critical role in the formation of the
hematopoietic microenvironment. In addition to providing the scaffolding (stromal)
fraction of the bone marrow on which hematopoietic stem cells proliferate,
mesenchymal stem cells are thought to play a role in hematopoiesis itself.77
While MSCs were initially identified in adult bone marrow, they have also been
isolated from fetal hematopoietic tissues where they attend to the migration of
hematopoietic development. Their precise identity remains poorly defined because
of the lack of specific markers. The ability of MSCs to self-renew and differentiate
into tissues of mesodermal origin (osteocytes, adipocytes, chondrocytes) and the
lack of expression of hematopoietic molecules are currently the focal criteria for
isolation.
Recently, mesenchymal stem cells have been shown to exert a profound
immunosuppressive effect on polyclonal as well as antigen-specific T cell responses
by inducing a state of division arrest anergy. Thus, the multi-potent capacity of
MSCs, their role in supporting hematopoiesis, and their immunoregulatory activity
make them particularly attractive for therapeutic exploitation.
For example, mesenchymal stem cells have been shown to significantly improve
hematopoietic recovery in patients receiving high-dose chemotherapy when
compared to autologous blood stem cell transfusion alone. When culture-expanded
MSCs are co-infused with autologous blood stem cells in breast cancer patients,
accelerated hematopoietic recovery is observed. 78
77 Dazzi F, Ramasamy R, Glennie S, Jones SP, Roberts I. The role of mesenchymal stem cells in haemopoiesis. Blood Rev 2006; 20: 161-171. 78 Koc ON, Gerson SL, Cooper BW, Dyhouse SM, Haynesworth SE, Caplan AI , et al. Rapid hematopoietic recovery after coinfusion of autologous-blood stem cells and culture-expanded marrow mesenchymal stem cells in advanced breast cancer patients receiving high-dose chemotherapy. J Clin Oncol 2000; 18: 307-316.
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C. Gene Transfer Studies
Mesenchymal stem cells have also been proposed for use in combinatorial gene
therapy protocols for the treatment of disease and the promotion of repair. The
efficacy of such a therapeutic approach depends on determining which vectors give
maximal transgene expression with minimal cell death. For this reason, numerous
studies are being carried out to determine vectors that can be used with minimum
danger and optimal effectiveness.
In a comparative study of viral and non-viral vectors for gene transfer into rat
mesenchymal stem cells, gene transfer via adenovirus, adeno-associated virus (AAV;
serotypes 1, 2, 4, 5, and 6), lentivirus, and nonviral vectors were compared.
Lentivirus proved to be most effective, with transduction efficiencies of up to 95%,
concurrent with low levels of cell toxicity.79
Gene therapy studies for specific treatment applications are also being conducted.
For instance, strategies using MSC-mediated gene therapy have been developed to
promote bone formation, through use of a modified adenoviral vector to introduce
the BMP2 gene.80 Potential applications of this strategy include treatment of
congenital bone defects, repair of extreme traumatic injury, and improvement of
growth velocity in children with osteogenesis imperfecta, a genetic disease
characterized by defective connective tissue (sometimes called “brittle bone
disease”).
Therapeutic benefits of gene-modified human MSCs have also been explored for the
ability to aid recovery after severe brain damage. Since angiogenesis is thought to be
of critical importance in repair of cerebral ischemia, Toyama, et al. generated gene-
modified human MSCs transfected with the angiopoietin-1 gene and VEGF gene, and
transplanted the cells into rats in which middle cerebral artery occlusion (MCAO)
79 McMahon JM, et al. Gene Transfer into Rat Mesenchymal Stem Cells: A Comparative Study of Viral and Nonviral Vectors. Stem Cells and Development 2006. 80 Tsuda H, et al. Efficient BMP2 gene transfer and bone formation of mesenchymal stem cells by a fiber-mutant adenoviral vector. Mol Ther 2003; 7(3): 354-365.
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had been induced.81 Based on comparison to test subjects that received hMSCs
lacking gene modification, it was concluded that localized Ang-1 and VEGF gene
expression assists in functional recovery after cerebral infarction in a rat model.
Gene transfer of MSCs could eventually be used to treat a diverse range of diseases
that involve failed cellular function, such as diabetes, stroke, congestive heart
failure, muscular dystrophy, and more.
D. Cell Transplantation for Site-Specific Repair
1. Overview
As mentioned already, MSCs can be induced to differentiate and express the
traits of a diverse range of cell types, including muscle, bone, skin, cartilage,
tendons, ligaments, marrow stroma, fat, neural cells, and more. For this reason,
MSCs represent a population of cells with the potential to contribute to
treatments for a diverse range of acute and degenerative diseases in which
tissues need regeneration or repair.
While mesenchymal stem cells have been explored for potential benefits in 223
different medical conditions,82 there are four research applications that
represent the majority of investigation to date. These areas are:
Osteogenic Repair
Myogenic and Myocardial Repair
Neural Repair
Pancreatic Repair
81 Toyama K, et al. Therapeutic benefits of angiogenetic gene-modified human mesenchymal stem cells after cerebral ischemia. Exp Neurol 2009; 216(1): 47-55. 82 “Deep Web” is a term that refers to general use Surface Web content, as well as content that is not indexed by standard search engines. The Deep Web contains over 900 billion pages, while the Surface Web is composed of only 25 billion pages. BioInformant’s Deep Web search allows access to unlimited search results (beyond the 50 page limit imposed by Google.com per search query), to screen text within web-based PDF files using image functionality, and to filter through large-scale public databases that include patent, scholar, grant, business, and university databases. Deep Web search query used: [“Mesenchymal Stem Cell” + {Database of Medical Conditions}]. Source of {Database of Medical Conditions}: www.diseasesdatabase.com. Query Date: May 1, 2013.
29
In each of these applications, MSCs represent an advantageous cell type for
allogenic transplant; as evidence indicates, MSCs are immune-privileged, with
low MHC I and no MHC II expression, which reduces risk of rejection and
complication during transplantation. 83
2. Major Applications
a. Osteochondral Repair
Mesenchymal stem cells are currently being investigated for use in the
treatment of a number of skeletal conditions. The osteogenic potential of
MSCs has been utilized to treat defective fracture healing, both alone and in
combination with scaffolds, in the repair of large bone defects.84 This
approach has been met with a high degree of success. MSCs have also been
used for cartilage repair. In a study conducted by Wakitani and colleagues,
autologous MSCs were expanded ex vivo, embedded in a collagen gel, and re-
implanted into areas of articular cartilage defect in osteoarthritis patients.85
It was concluded that formation of hyaline cartilage-like tissue was improved
in the experimental group versus the control group.
Although most applications for tissue repair involve local transplantation of
MSCs to directly target an area of injury, systemic transplantation of MSCs
has been in place for a long time in the form of hematopoietic stem cell
transplants. Successful application of systemic MSC transplant has been
performed in children with osteogenesis imperfecta. In a study conducted by
Horwitz and colleagues, children with osteogenesis imperfecta received
systemic infusion of allogenic MSCs. Transplanted MSCs migrated to bone
83 Uccelli A, Moretta L, Pistoia V. Immunoregulatory function of mesenchymal stem cells. Eur J Immunol 2006; 36: 2566-2573. 84 Quarto R, Mastrogiacomo M, Cancedda R, Kutepov SM, Mukhachev V, Lavroukov A , et al. Repair of large bone defects with the use of autologous bone marrow stromal cells. N Engl J Med 2001; 344: 385-386. 85 Wakitani S, Imoto K, Yamamoto T, Saito M, Murata N, Yoneda M. Human autologous culture expanded bone marrow mesenchymal cell transplantation for repair of cartilage defects in osteoarthritic knees. Osteoarthritis Cartilage 2002; 10: 199-206.
30
and produced collagen, thus suggesting that mesenchymal stem cells may
represent a novel approach for treating this debilitating genetic condition.86
Applications of mesenchymal stem cells in orthopedics are on the rise.
Currently, the interdisciplinary orthopedics market is sized at US $100 million
annually, but it is expected to surpass the $3 billion mark within the next
decade.87 As orthopedic problems ranging from back pain to osteoporosis
plague 75 million Americans, or approximately a quarter of the US
population, it is apparent that the demand for MSC-based orthopedic
treatments will continue to rise. Bio-pharmaceutical88 and pharma
companies will consider development of MSC-based orthopedic therapies a
priority area for research and development due to the high valuation of the
orthopedics market and its accelerating growth.
b. Myogenic and Myocardial Repair
A number of groups have reported mesenchymal stem cell differentiation
into cardiomyocytes in vitro. The current in vivo approach consists of
injecting undifferentiated MSCs or whole bone marrow directly into the
heart. Although the underlying mechanisms remain to be understood,
significant improvement has been observed, suggesting that MSC infusion
triggers the formation of new cardiomyocytes and neoangiogenesis in the
human heart.89,90
However, it is still unclear whether MSCs act directly by in situ differentiation
or fusion with resident myocytes91, or indirectly through secretion of pro-
86 Horwitz EM, Prockop DJ, Fitzpatrick LA, Koo WW, Gordon PL, Neel M , et al. Transplantability and therapeutic effects of bone marrow-derived mesenchymal cells in children with osteogenesis imperfecta. Nat Med 1999; 5: 309-313. 87 Stem Cells & Orthopedics - Brown University – 2008 – BioMed Division. Available at: http://biomed.brown.edu/Courses/BI108/BI108_2008_Groups/group05/Orthopedics.html [Accessed May 10,2013]. 88 Biopharmaceutical products are pharmaceuticals derived from life forms. 89 Wollert KC, Meyer GP, Lotz J, Ringes-Lichtenberg S, Lippolt P, Breidenbach C, et al. Intracoronary autologous bone-marrow cell transfer after myocardial infarction: The BOOST randomized controlled clinical trial. Lancet 2004; 364: 141-148. 90 Fuchs S, Satler LF, Kornowski R, Okubagzi P, Weisz G, Baffour R, et al. Catheter-based autologous bone marrow myocardial injection in no-option patients with advanced coronary artery disease: A feasibility study. J Am Coll Cardiol 2003; 41: 1721-1724. 91 Lee JH, Kosinski PA, Kemp DM. Contribution of human bone marrow stem cells to individual skeletal myotubes followed by
myogenic gene activation. Exp Cell Res 2005; 307: 174-182.
31
myogenic factors promoting endogenous myocardial repair, such as VEGF
and FGF.92
c. Neural Repair
The ability of mesenchymal stem cells to migrate to the site of injury has also
been reported following transplantation in the brain. MSCs transplanted into
rat striata were observed to migrate across the corpus callosum and
populate the striatum, thalamic nuclei, and substantia nigra of a lesioned
hemisphere.93 Untreated MSCs systemically infused into animals with
damaged brain tissue have also been seen to migrate to the trauma site and
improve recovery, although whether this is via secretion of neuroprotective
factors or by differentiation into neural tissue remains unclear.
While it is not disputed that the MSCs appear to serve a positive role in
recovery, there is debate as to whether the signs of differentiation observed
in situ are real or simply a result of cell fusion with resident neural cells.
More research is needed to determine the precise extent of MSC
contribution in brain repair models, since such results could have
implications for neurodegenerative diseases such as Parkinson's disease and
Alzheimer's disease, and traumatic events such as stroke or spinal cord
injury.
Although the adult brain contains populations of neural stem cells, these are
insufficient to replace the massive number of cells required to treat these
conditions on a functional level.94 To date, the critical difficulty with treating
neurological degeneration, brain defects, and head trauma has been the
difficulty of sourcing neural progenitor cells acceptable for use in
transplantation. While fetal tissue and differentiated embryonic stem cells
92 Xu M, Uemura R, Dai Y, Wang Y, Pasha Z, Ashraf M. In vitro and in vivo effects of bone marrow stem cells on cardiac structure and function. J Mol Cell Cardiol 2006. 93 Hellmann MA, Panet H, Barhum Y, Melamed E, Offen D. Increased survival and migration of engrafted mesenchymal bone marrow stem cells in 6-hydroxydopamine-lesioned rodents. Neurosci Lett 2006; 395: 124-128. 94 Gage FH. Mammalian neural stem cells. Science 2000; 287: 1433-1438.
32
have been considered for this purpose, these sources suffer limitations that
included limited tissue availability and ethical and safety concerns.
The identification of an adult population of cells, such as mesenchymal stem
cells, which can be easily obtained from autologous or donated marrow and
can be cultured and manipulated ex vivo, would represent a significant
breakthrough in the search for many applications in neuro-regenerative
medicine. Because clinical trials impacting the brain are particularly
dangerous to perform in humans, to date, most of this research has been
performed in animal models.
d. Pancreatic Repair
As mentioned, it has also been demonstrated that MSCs have the potential
to differentiate into beta-pancreatic islet cells.95 Specifically, Chen and
colleagues demonstrated differentiation of rat bone marrow-derived
mesenchymal stem cells in vitro into functional islet-like cells. This finding
represents potential use of MSCs to treat a range of pancreatic-related
diseases, including:
Diabetes
Pancreatic cancer
Cystic fibrosis-induced damage of the pancreas (debilitating)
Congenital malformations of the pancreas
Currently, the US Center for Disease Control (CDC) estimates that
approximately 24 million adults in USA have diabetes, mostly type-2 diabetes
that is associated with a poor diet and lack of exercise. More frightening,
however, is that the CDC projects that a third of the US population will have
95 Chen LB, Jiang XB, Yang L. Differentiation of rat marrow mesenchymal stem cells into pancreatic islet beta-cells. World J of Gastroenterol 2004; 10(20): 3016–3020.
33
diabetes by 2050.96 Clearly, MSC-based treatments for diabetes will be a
major medical priority for the next several decades.
E. Topical Therapy and Use of MSCs in Wound Healing
Another application for which mesenchymal stem cells have demonstrated utility is
in topical therapy involving the treatment of wounds, or ischemic or damaged tissue.
It has been noted in several studies that characteristics of MSCs are activated upon
contact with injured sites.97,98 While MSCs can be administered systemically for
homing to target sites, vascular transit risks that MSCs will be taken out of
circulation, on either a temporary or a permanent basis, in organs such as the lungs,
spleen, and liver.99 This can significantly reduce the numbers of cells that reach
target sites, where they must exit the vasculature and enter the connective tissue
stroma to exert a therapeutic effect. Thus, direct application of mesenchymal stem
cells to wound sites appears to be preferable.
While interaction of MSCs with other wound cells through paracrine mechanisms
are not clearly understood, interactions with vascular endothelial cells and
immunomodulation appear to play significant roles in accelerating wound healing
and in reducing scar formation upon the completion of the healing process.100
A distinguishing factor for use of MSCs in topical wound healing is the mechanism by
which the delivery of these cells is accomplished. Two different approaches for
topical delivery are summarized below:
96 National Diabetes Fact Sheet - Center for Disease Control (CDC). Available at: http://www.cdc.gov/diabetes/statistics/index.htm [Accessed May 11, 2013]. 97 Caplan AI, Dennis JE. Mesenchymal stem cells as trophic mediators. J Cell Biochem 2006; 98: 1076-1084. 98 Caplan AI. Why are MSCs therapeutic? New data: New insight. J Pathol 2009; 217: 318-324. 99 Karp JM, Teo GSL. Mesenchymal stem cell homing: The devil is in the details. Cell Stem Cell 2009; 4: 206-216. 100 Sorrell JM, Caplan AI. Topical delivery of mesenchymal stem cells and their function in wounds. Stem Cell Research & Therapy 2010; 1: 30.
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1. Stoff and colleagues tested injection of concentrated human MSCs (hMSCs)
adjacent to incisional wounds in the skin of rabbits.101 The human MSCs were
observed to traverse the dermal-epidermal junction region of the wound within
two weeks and reach the margin between the wound bed and the underlying
fascia within three weeks. Interestingly, the wounds treated with hMSCs
recovered 52% of the normal tensile strength of skin, compared to only a 31%
recovery of tensile strength for non-treated wounds. This difference was likely a
result of the hMSC treated wounds presenting with fuller and more organized
deposition of collagenous fibers.
2. Falanga and colleagues chose to approach wound repair by incorporating
autologous MSCs into a fibrin spray for topical delivery.102 Using an experimental
mouse model, the researchers made excisional wounds into the skin on the tail
of genetically diabetic (db/db) mice. They then administered the fibrin spay with
and without the presence of autologous MSCs. Interestingly, wounds treated
with the MSCs healed more quickly and presented with a more mature histology
than wounds in which MSCs were not applied. Using autologous MSCs, Falanga
and colleagues subsequently performed a parallel study on human chronic non-
healing wounds and observed similar outcomes in accelerated wound-healing for
MSC-treated sites.
In addition to these two approaches, another interesting (theoretical) approach is
direct application of MSCs to skin wounds through integration of MSCs into skin
equivalents, using bilayers composed of an epidermis overlying a fibroblast-based
dermis.103 Because mesenchymal stem cells are a fibroblastic population, they can
be used alone or paired with dermal fibroblasts to form the dermal component of
skin equivalents.104 Incorporation of MSCs into a fibroblast matrix has been
101 Stoff NS, Moore ST, Numnum M, Espinosa-de-los-Monteros A, Richter DF, Siegal GP, Chow LT, Feldman D, Vasconez LO, Mathis JM, Stoff A, Rivera A, Banerjee A, Stoff-Khalili MA, Curiel DT. Promotion of incisional wound repair by human mesenchymal stem cell transplantation. Exp Dermatol 2009; 18: 362-369. 102 Falanga V, Iwamoto S, Chartier M, Yufit T, Butmarc J, Kouttab N, Shrayer D, Carson P. Autologous bone marrow-derived cultured mesenchymal stem cells delivered in a fibrin spray accelerate healing in mouse and human cutaneous wounds. Tissue Eng 2007; 13: 1299-1312. 103 Boyce ST. Cultured skin substitutes: A review. Tissue Eng 1996; 2: 255-266. 104 Fioretti F, Lebreton-DeCoster C, Gueniche F, Yousfi M, Humbert P, Godeau G, Senni K, Desmoulière A, Coulomb B. Human bone marrow-derived cells: An attractive source to populate dermal substitutes. Wound Repair Regen 2008; 16: 87-94.
35
demonstrated to improve the angiogenic potential of that matrix,105 which suggests
that such a system might also be capable of supporting accelerated site repair.
F. Use of 3-D Scaffolds for Tissue Engineering
To mimic the three-dimensional nature of mesenchymal stem cell derivatives (bone,
cartilage, muscle, and fat), researchers now recognize that it may be important to
incorporate 3-D scaffolding when exploring MSC differentiation in vitro. Collagen, a
normal constituent of bone, is one candidate being used as a substrate on which to
engineer bone and cartilage from mesenchymal-derived precursors. In a recent
study, a collagen-glycosaminoglycan scaffold was used to provide a suitable 3-D
environment on which to culture adult rat mesenchymal stem cells and induce
differentiation along osteogenic and chondrogenic lineages.106
The results demonstrated that adult rat mesenchymal stem cells can undergo
osteogenesis when grown on the collagen-glycosaminoglycan scaffold and
stimulated with osteogenic factors, as evaluated by the temporal induction of the
bone-specific proteins, collagen I and osteocalcin, and matrix mineralization.
As well as supporting osteogenesis, when the cell-seeded scaffold was exposed to
chondrogenic factors, collagen II immunoreactivity was increased, providing
evidence that the scaffold can also provide a suitable 3-D environment that supports
chondrogenesis. 107
Due to promising results in this area that have established that MSCs differentiate
more favorably in 3-D scaffolds, more research of this type will continue to be
performed in both human and non-human species models.
105 Sorrell JM, Baber MA, Caplan AI. Influence of adult mesenchymal stem cells on in vitro vascular formation. Tissue Eng Part A 2009; 15: 1751-1761. 106 Ferrell, et al. A Collagen-glycosaminoglycan Scaffold Supports Adult Rat Mesenchymal Stem Cell Differentiation Along Osteogenic and Chondrogenic Routes. Tissue Engineering 2006. 107 Ferrell, et al. A Collagen-glycosaminoglycan Scaffold Supports Adult Rat Mesenchymal Stem Cell Differentiation Along Osteogenic and Chondrogenic Routes. Tissue Engineering 2006.
36
G. Drug Delivery Applications: Engineered MSCs as specific carriers of anti-cancer drugs
Another interesting application for MSCs is their potential use in drug delivery
applications. It is generally hypothesized that a pool of reserve mesenchymal stems,
involved in tissue homeostasis, may be supported by circulating bone marrow MSCs,
as shown by injecting the reserve MSCs intravenously: these cells spread into all
tissues, but preferentially survive and multiply in the presence of regenerating
tissues and tumors, where they become vasculo-stromal fibroblasts.108 For this
reason, engineered MSCs may have utility as specific carriers of anti-cancer drugs,
since circulatory infusion of MSCs appears to be feasible and safe for short
durations.109
Theoretically, MSCs represent an ideal system for tumor-localized delivery of anti-
cancer agents, as the cells possess tumor-tropic properties and at the same time are
compatible and non-immunogenic to the host.110
For instance, Houghton, et al.111 reported MSC engraftment into gastric glands in a
model of gastric cancer. Khakoo, et al.112 demonstrated that human MSCs injected
intravenously localize to sites of tumorigenesis in a model of Kaposi's sarcoma.
Studies have shown that injection of MSCs in the contralateral hemisphere, the
carotid vein, or the tail vein leads to MSC location in a tumor in the other
hemisphere.113,114 These examples show that MSCs exhibit tropism to tumor
microenvironments.
108 Krampera M, Pizzolo G, Aprili G, Franchini M. Mesenchymal stem cells for bone, cartilage, tendon and skeletal muscle repair. Bone 2006; 39: 678–683. 109 Devine SM, Bartholomew AM, Mahmud N, Nelson M, Patil S, Hardy W, et al. Mesenchymal stem cells are capable of homing to the bone marrow of non-human primates following systemic infusion. Exp Hematol 2001; 29: 244–255. 110 Mesenchymal stromal cells as a drug delivery system - Menon, Shi, Carroll - Brain Tumor Research Laboratory - Department of Neurosurgery - Brigham & Women's Hospital. Available at: http://www.stembook.org/node/534#sec1-3 [Accessed May 12, 2013]. 111 Houghton J, Stoicov C, Nomura S, Rogers AB, Carlson J, Li H, Cai X, Fox JG, Goldenring JR, Wang TC. Gastric cancer originating from bone marrow-derived cells. Science 2004; 306: 1568–1571. 112 Khakoo AY, Pati S, Anderson SA, Reid W, Elshal MF, Rovira II, Nguyen AT, Malide D, Combs CA, Hall G, et al. Human mesenchymal stem cells exert potent antitumorigenic effects in a model of Kaposi's sarcoma. J Exp Med 2006; 203: 1235–1247. 113 Nakamura K, Ito Y, Kawano Y, Kurozumi K, Kobune M, Tsuda H, Bizen A, Honmou O, Niitsu Y, Hamada H. Antitumor effect of genetically engineered mesenchymal stem cells in a rat glioma model. Gene Ther 2004; 11: 1155–1164. 114 Nakamizo A, Marini F, Amano T, Khan A, Studeny M, Gumin J, Chen J, Hentschel S, Vecil G, Dembinski J et al. Human bone marrow-derived mesenchymal stem cells in the treatment of gliomas. Cancer Res 2005; 65: 3307–3318.
37
Unfortunately, this area of MSC application is not without flaw. MSCs also exhibit
tropism to sites of wounds, ischemic or damaged tissue, and chronic inflammation,
all of which can complicate targeted delivery of a cancer-specific therapeutic agent.
H. Drug Screening
In addition, mesenchymal stem cells may eventually have the potential to assist in
drug development, as they allow for earlier identification of adverse side effects in
new drug candidates. Currently, new drug prospects go through animal testing
before approval for use in human trials. However, animal models do not identically
replicate the physiological conditions of the human response, and consequently, it is
possible for drugs that pass safety testing in animals to produce severe effects in
humans. An appropriate solution to this would be to test new drug prospects on
human cells before allowing them to be administered in human clinical trials.
Because most harmful side effects caused by drug candidates are complications with
the liver, kidney, or heart, the possibility of differentiating MSCs into hepatocytes,
renal cells, or cardiac myocytes for use in early stage drug toxicity screening would
represent an application with extremely high industry demand.
The theory behind drug toxicity testing on human stem cell-derived populations is
that drug companies could collect populations of stem cells from individuals with a
wide variety of genetic backgrounds. In addition to allowing for effective and specific
toxicity testing, this approach would also assist with early identification of whether
genetic populations do or do not respond well to a drug candidate, a novel approach
to personalized medicine.
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I. MSC-Mediated Inhibition of Immune Effector Cells
MSCs are believed to be immunosuppressive through a mechanism thought to
involve paracrine inhibition of T- and B-cell proliferation,115,116 and as such have also
been used in trials investigating their effects on autoimmune diseases and Graft
Versus Host Disease (GVHD). Co-infusion of donor-derived MSCs together with
hematopoietic stem cells (HSCs) can reduce the incidence and severity of GVHD in
sibling allografts.117 The hypo-immunogenic properties of MSCs are considered by
some to be sufficient to allow transplantation even between individuals who are not
HLA-compatible.118
Because allogenic transplantation studies are somewhat difficult to carry out on
human patients, animal models are frequently used as an alternative.
In addition, osteoarthritis (OA) is the most common arthritic condition experienced
in human populations, but like rheumatoid arthritis (RA), it presents as an
inflammatory environment with immunological involvement. To date, this has
presented a major obstacle with regard to traditional cartilage tissue engineering
approaches for use in treatment of OA. However, recent advances in the
understanding of MSCs have demonstrated that MSCs possess potent
immunosuppression and anti-inflammation effects. Through secretion of various
soluble factors, MSCs can influence the local tissue environment and exert
protective effects with an end result of effectively stimulating regeneration in situ.119
This function of MSCs is beginning to be explored for use in the treatment of
degenerative joint diseases that include osteoarthritis and rheumatoid arthritis.
115 Di Nicola M, Carlo-Stella C, Magni M, Milanesi M, Longoni PD, Matteucci P, et al . Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood 2002; 99: 3838-3843. 116 Uccelli A, Moretta L, Pistoia V. Immunoregulatory function of mesenchymal stem cells. Eur J Immunol 2006; 36: 2566-2573. 117 Lazarus HM, Koc ON, Devine SM, Curtin P, Maziarz RT, Holland HK , et al. Cotransplantation of HLA-identical sibling culture-expanded mesenchymal stem cells and hematopoietic stem cells in hematologic malignancy patients. Biol Blood Marrow Transplant 2005; 11: 389-398. 118 Le Blanc K, Tammik C, Rosendahl K, Zetterberg E, Ringden O. HLA expression and immunologic properties of differentiated and undifferentiated mesenchymal stem cells. Exp Hematol 2003; 31: 890-896. 119 Chen F, Tuan R. Mesenchymal stem cells in arthritic diseases. Arthritis Research & Therapy 2008; 10: 223.
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VI. APPLICATION PRIORITIES
A. Overall
Considering the applications described above, the application receiving the most
attention across the research community is the use of MSCs in transplantation for
site-specific repair, particularly osteochondral and myocardial repair.
A breakdown of research frequency for the most common applications is presented
below:
TABLE: Breakdown of MSC Research Rates by Application (January 1, 2012 – January 1, 2013)120
MSC APPLICATION % OF RESEARCH ACTIVITY
Cell Transplantation for Site-Specific Repair 23.4
Basic Research 22.1
Drug Screening 13.7
Tissue Engineering 12.3
Drug Delivery 9.7
Gene Therapy 9.1
Supporting Hematopoietic Cells 3.0
Inhibition of Immune Effector Cells 1.1
Topical Therapy 0.9
Other 4.7
TOTAL 100.0
120 Hybridized Statistical Analysis. Data Input Sources: 1) Publication rate data from PubMed (http://www.ncbi.nlm.nih.gov/pubmed/); 2) Publication rate data from Highwire Press (http://highwire.stanford.edu/); 3) Patent rate data from USPTO (http://www.uspto.gov/); 4) Web query rates for: “Mesenchymal Stem Cell” + [Database of Product Terms]. Test Interval: Jan 1, 2012 – Jan 1, 2013.
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B. By Segment
1. Academic
Within the academic community, basic research applications of MSCs is the
dominant research activity, although this interest in understanding cellular
mechanisms by no means diminishes the pursuit of clinical applications by
academic researchers. MSCs simply represent a complicated and highly valuable
cell type, and the academic sector is one segment of the research community
that has freedom to explore fundamental principles governing this cell type
without needing to link investigation to immediate commercial benefit.
The following are areas of basic research frequently being explored within the
academic research community:
MSC Morphology: This area is relatively well understood due to a variety
of imaging techniques used to characterize MSC morphology since their
discovery in the 1960s.
23.4%
22.1%
13.7%
12.3%
9.7%
9.1%
3.0% 1.1% 0.9%
4.7%
Breakdown of MSC Research Rates by Application
(January 1, 2012 – January 1, 2013)
Cell Transplantation for Site-Specific Repair
Basic Research
Drug Screening
Tissue Engineering
Drug Delivery
Gene Therapy
Supporting Hematopoietic Cells
Inhibition of Immune Effector Cells
Topical Therapy
Other
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MSC Detection: Currently, a definitive test does not exist to determine
whether a single cell is a mesenchymal stem cell. While surface antigens
can be used to isolate a population of cells that have comparable self-
renewal and differentiation capacities, MSCs as a population do not
usually express all of the same biological markers. Furthermore, there is
still uncertainty as to which markers should be expressed in order to
classify a cell as an MSC. Because of the heterogeneous nature of MSC
populations, it may be that the therapeutic properties attributed to MSCs
are a consequence of interactions among the different cells that compose
an MSC culture, which would suggest that there is no one cell that has all
the properties.
Because this is a complicated and controversial area of basic MSC
research, it has received a significant amount of attention within the past
few years and will continue to remain an interesting area of MSC
research.
Differentiation Capacity: MSCs demonstrate great capacity for self-
renewal while maintaining multi-potency. Beyond that, there is little that
can be definitively said about their differentiation capacity. The standard
test to confirm MSC multi-potency is to differentiate the cells into their
distinct lineages of osteoblasts, adipocytes, and chondrocytes.
However, it is critical to note that the degree to which an MSC culture will
differentiate varies both by tissue source of origin and by donor. At this
time, it is not clear whether this variation is due to varying amounts of
"true" progenitor cells present within cultures or variable differentiation
capacities of progenitor cells from person to person. For example, it is
known that the capacity of MSCs to proliferate and differentiate
decreases with the age of the donor, as well as the time in culture, but it
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is not known whether this is due to a decrease in the number of MSCs or
physiological change within existing MSCs.
Immuno-modulatory Effects: Many studies have indicated that human
MSCs avoid allorecognition, interfere with dendritic cell and T-cell
function, and generate a local immunosuppressive microenvironment by
secreting cytokines.121 It has also been shown that the immuno-
modulatory function of human MSC is enhanced when the cells are
exposed to an inflammatory environment characterized by the presence
of elevated local interferon-gamma levels.122 Other studies disagree with
some of these findings, reflecting both the highly variable nature of MSC
isolates and the substantial differences between isolates generated by
the numerous methods under development. Contradictory findings in
this area will continue to generate research attention.
2. Biotechnology
Within the biotechnology community, exploring clinical applications of
mesenchymal stem cell is the dominant focus, with special attention given to
osteogenic and cardiovascular applications.
Within the US, the leading biotech company dealing with clinical applications of
MSCs is Osiris Therapeutics, Inc. (symbol: OSIR), which already has a range of
products in development and on the market, including one called Osteocel.
Osteocel is an unexpanded preparation of mesenchymal stem cells extracted
from bone marrow aspirate. Osiris also has products for graft-versus-host
disease (GVHD), cartilage applications, and cardiac applications. Osiris currently
has a market cap of over $400 million.123
121 Ryan JM, Barry FP, Murphy JM, Mahon BP. Mesenchymal stem cells avoid allogeneic rejection. J Inflamm (Lond) 2005; 2: 8. 122 Ryan JM, Barry F, Murphy JM, Mahon BP. Interferon-gamma does not break, but promotes the immunosuppressive capacity of adult human mesenchymal stem cells. Clin Exp Immunol 2007; 149(2): 353–363. 123 Osirius Pharmaceuticals, Securities and Exchange Commission (SEC) Filing, 4th Quarter 2011, Form 10Q.
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Worldwide, the leading company developing MSC therapies is an Australia-based
company called Mesoblast. Mesoblast has a US division called Angioblast.
Mesoblast's core technology is based on the identification, extraction, and
enrichment of mesenchymal precursor cells (MPCs).124 Mesoblast has two Phase
IIa trials focused on spinal fusion and cardiovascular disease, has completed pilot
studies in humans in long bone fractures and early stage intervertebral disc
cartilage, and is completing large animal studies in knee cartilage and
osteoarthritis. The spinal fusion trial is of an allogeneic product for patients with
severe intervertebral disc disease requiring spinal fusion. Allied to this trial is a
preclinical trial for repair and regeneration of vertebral disc cartilage.
Mesoblast’s US division, Angioblast, recently completed a pilot clinical trial of an
autologous MSC product at John Hunter Hospital designed to treat cardiac
damage. Angioblast is also doing a Phase IIa trial in Texas using allogeneic MSCs.
Both Mesoblast and Angioblast work closely with manufacturers Cordis and
Biosense (J&J companies), as well Medtronic, which makes the biomedical
support matrix used for delivery of the cells in orthopedic applications.
In addition, on December 8, 2010, Mesoblast entered into a strategic
collaboration with Cephalon, Inc., a US-based pharmaceutical company, to
develop and commercialize adult Mesenchymal Precursor Stem Cell (MPC)
therapies for degenerative conditions of the cardiovascular and central nervous
systems.125 These conditions include: Congestive heart failure, Acute myocardial
infarction, Parkinson's disease, and Alzheimer's disease. The agreement also
includes products for augmenting hematopoietic stem cell transplantation in
cancer patients.
Within the agreement, in exchange for exclusive world-wide rights to
commercialize specific products based on Mesoblast's unique adult stem cell
technology platform, Cephalon is providing an up-front sum to Mesoblast of US
124 Mesoblast Corporate Website. Available at: http://www.mesoblast.com/. [Accessed March 15, 2012]. 125 Cephalon, Mesoblast to develop, commercialize novel adult MPC therapeutics – MedicalNews.com. Available at: http://www.news-medical.net/news/20101208/Cephalon-Mesoblast-to-develop-commercialize-novel-adult-MPC-therapeutics.aspx [Accessed March 15, 2012].
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$130 million, and has agreed to make regulatory milestone payments of up US
$1.7 billion, depending on clinical trial outcomes.
Mesoblast has agreed to be responsible for the expenses of certain Phase IIa
clinical trials and commercial supply of the products, while Cephalon will be the
funding source for all Phase IIb and III clinical trials, as well as resulting
commercialization of the products. Mesoblast will retain all manufacturing rights
and will share significantly in the net product sales. In addition, Cephalon is
purchasing a 19.99% stake in Mesoblast at a price of approximately US $220
million.
Clearly, the decision of Cephalon, Inc., to spend over US $130 million in an up-
front payment, as well as $200 million toward the purchase of a Mesoblast stock
position, suggests that the company strongly believes in the therapeutic viability
and the commercial market potential for adult Mesenchymal Precursor Stem Cell
(MPC) therapies.
3. Pharmaceutical
Because most biotech companies exploring clinical applications of mesenchymal
stem cells need outside funding to support their clinical trials, biotechnology and
pharmaceutical companies often demonstrate aligned priorities with regard to
MSC applications, as demonstrated by the Mesoblast and Cephalon alliance.
Similar to biotech companies, the dominant priority of pharma companies is to
explore clinical applications of mesenchymal stem cells for treating osteogenic
conditions (particularly bone repair and osteoarthritis) and cardiac damage
(particularly cardiac infarction and ischemia).
Baxter, Inc., an Illinois-based pharmaceutical company, treated their first patient
with blood-derived stem cells as part of a national Phase II trial in 2008. The
study aimed to alleviate the effects of chronic myocardial ischemia and used
45
MSCs harvested from the blood of each patient. Worldwide, several other
studies of similar focus are being designed as well.
Because of their tumor-trophic properties, there is also significant interest within
the pharmaceutical sector in genetic manipulation of MSCs, including the ability
to over-express target receptors (designed to improve site-specific honing
properties) and the introduction of exogenous genes (for purposes of secreting
active therapeutic agents upon site-specific localization). Any genes that could
exert antiproliferative or pro-apoptotic behavior upon localization of MSC-
expressing cells to sites of cancer could have utility for this purpose.
Interestingly, it was also recently recognized that in addition to localizing to sites
of solid tumors, mesenchymal stem cells are involved in regulation of
haematological malignancies,126 suggesting further potential for therapeutic
application of MSCs in the control of malignant conditions.
126 Stagg J. Mesenchymal Stem Cells in Cancer. Stem Cell Rev 2008; 4(2):119-124.
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VII. COMPANIES OFFERING HUMAN MSC RESEARCH PRODUCTS
As mentioned earlier, MSCs are a well-characterized population of adult stem cells,
regarding which 20,673 scientific articles have been published.127 Not surprisingly, this
level of research activity has attracted the attention of research supply companies, and
a substantial number of MSC research products now exist. There are currently 16
companies that offer human mesenchymal stem cell products and eight that offer rat
mesenchymal stem cell products.
Market competitors that offer human MSC research products are presented below, in
alphabetical order:
A. Abcam (www.abcam.com)
Abcam, located in Cambridge, MA, supplies a wide range of antibodies to cell surface
markers of human, mouse, and rat mesenchymal stem cells. Currently, the company
has 46 products categorized under its “Mesenchymal Stem Cells” product heading.
Of those, 40 are primary antibody products, four are proteins and peptides, and two
are kits.128
The kits are “Mouse Mesenchymal Stromal Cell Marker Antibody Panels” designed
for characterization of mouse multi-potent mesenchymal stromal cells. They contain
a group of antibodies for positive and negative selection of this lineage. One kit
provides CD90, CD29, CD44, and Sca-1 for positive selection and CD45 for negative
selection. The other kit provides CD105, CD90, CD44, and CD29 for positive selection
and CD45 for negative selection of the lineage. Thus, the kits vary only in inclusion of
Sca-1 versus CD105 for use in positive selection.
127 Figure calculated using search of Pubmed Database (http://www.ncbi.nlm.nih.gov/pubmed) for the search terms: ["Mesenchymal Stem Cell" OR "Mesenchymal Stem Cells"] OR ["Marrow Stromal Cell" OR "Marrow Stromal Cells"] OR ["Multi-potent Stromal Cell" OR "Multi-potent Stromal Cells"] OR ["Colony-Forming Unit-fibroblast" OR "Colony-Forming Unit-fibroblasts"]. Search encompassed four technical terms for the cell type, as well as variations in singular versus plural usage of terminology. . Executed March 15, 2012. 128 Mouse Mesenchymal Stromal Cell Marker Panel (CD44, CD90, Sca-1, CD45 and CD29) (ab93759) – Abcan, Corporate Website – Product page. Available at: http://www.abcam.com/Mouse-Mesenchymal-Stromal-Cell-Marker-Panel-CD44-CD90-Sca-1-CD45-and-CD29-ab93759.html [Accessed May 12, 2013].
47
B. BD Biosciences (www.bd.com)
BD Biosciences, headquartered in Franklin Lakes, NJ, offers a range of antibodies
reactive to mesenchymal stem cell markers. The company also offers a very
developed product line designed for general stem cell research, including matrices,
reagents, and extracellular matrix molecules.
BD Biosciences currently offers 696 products under the “Mesenchymal Stem Cells”
product heading, of which 25 are multi-color antibodies, 657 are single color/pure
antibodies, and 14 are magnetic cell separation products.129 The company also offers
90 documents and web pages that relate to the search term “mesenchymal stem
cells,” indicating the company is positioning itself as a resource for MSC related
information, attracting researchers to the site.
C. Celprogen
Celprogen also offers a sizeable selection of human MSC products, including stem
cell culture kits for differentiation, expansion, and maintenance of stem cells in an
undifferentiated state in a chemically defined extracellular matrix. Products that
Celprogen offers include:130
Human Mesenchymal Adipose Stem Cell Diff. Media with Serum (M36096-23DS )
Human Mesenchymal Bone Marrow Serum Free Differentiation Media (M36094-21D)
Human Mesenchymal Stem Cells derived from Human Cord Blood (36094-21)
Human Mesenchymal Stem Cells derived from Human Liver (36094-22)
Human Mesenchymal Stem Cells derived from Human Adipose Tissue (36094-23)
Additional related products
129 BD Biosciences, Corporate Website. Available at: www.bdbiosciences.com [Accessed May 11, 2013]. 130 Celprogen, Corporate Website. Available at: www.celprogen.com [Accessed May 10, 2013].
48
Interestingly, Celprogen does not appear on the first page of Google Search results
for any of the common phrases that would be used to search for their mesenchymal
stem cell products. For instance, using the search phrases “Mesenchymal Stem Cell”
and “Mesenchymal Stem Cells,” Celprogen does not even appear in the first 20
pages of Google Search Results.131 Therefore, most researchers looking for
mesenchymal stem cells would be unlikely to uncover Celprogen as a provider using
online search. As a result, even though Celprogen has a very well-developed
product line, at this time, the company is not well-positioned from a visibility
standpoint.
Currently, the company offers 124 products specific to mesenchymal stem cell
research.
D. Cyangen Biosciences (www.cyagen.com)
Cyagen Biosciences, a bioscience company located in the Science Park of
Guangzhou, China, provides human and animal stem cells, their genetically modified
derivatives, and culture and research reagents for use with these cell types. The
company’s 1800 square meter facility has a laminar flow ultra-clean cell culture lab
built to GMP standard, a molecular biology lab, and an animal barrier facility housing
mice and rats intended for transgenic and gene knockout research. At this time,
Cyagen is not yet a competitor with respect to mesenchymal stem cell products
within the United States, because the company only distributes products inside
China and does not ship across the border. However, Cyagen is in talks with US
companies about distribution of their products in North America and has plans to
enter into the North American market during 2011. Thus, Cyagen will be a direct
competitor when and if they establish a US distributor.
131 Google Web Search (www.google.com) for phrases, “Mesenchymal Stem Cell” and “Mesenchymal Stem Cells.” Executed May 5, 2013.
49
Cyagen currently offers 20 mesenchymal stem cell specific products, listed below:132
PRODUCT NAME
Mesenchymal Stem Cell Chondrogenic Differentiation Medium
Balb/c Mouse Mesenchymal Stem Cell
Mesenchymal Stem Cell Adipogenic Differentiation Medium
Mesenchymal Stem Cell Osteogenic Differentiation Medium
Dog Mesenchymal Stem Cell
Rabbit Mesenchymal Stem Cell
F344 Rat Mesenchymal Stem Cell
SD Rat Mesenchymal Stem Cell
Wistar Rat Mesenchymal Stem Cell
Cynomologus Monkey Mesenchymal Stem Cell
Rhesus Monkey Mesenchymal Stem Cell
Human Fetal Mesenchymal Stem Cell
Human Mesenchymal Stem Cell
Sprague-Dawley Rat Adipose-derived Mesenchymal Stem Cell
C57BL/6 Mouse Adipose-derived Mesenchymal Stem Cells
Human Adipose-derived Mesenchymal Stem Cell
C57BL/6 Mouse Mesenchymal Stem Cell
Dog Mesenchymal Stem Cells
Rabbit Mesenchymal Stem Cells
Sprague-Dawley Rat Mesenchymal Stem Cells
As of January 1, 2009, Cyagen offered only five MSC-specific research products,
meaning that in a little over three years, Cyagen has multiplied its MSC product
offerings four-fold. If this rate of product growth continues and the company aligns
itself with a global distributor, Cyagen will become force within the MSC research
products sector.
E. Life Technologies (www.lifetechnologies.com)
Invitrogen, located in Carlsbad, CA, also offers a wide range of mesenchymal stem
cell products. The company currently offers 159 products under its “Mesenchymal
Stem Cells” product title.133
Of those products:
76 are primary antibodies
54 are growth factors, chemokines, and cytokines
132 Cyagen Biosciences, Corporate Website. Available at: www.cyagen.com [Accessed May 10 , 2013]. 133 Invitrogen, Corporate Website. Available at: www.invitrogen.com [Accessed May 9, 2013].
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6 are serum-free and specialty media
4 are extracellular matrix proteins
The rest represent product categories with three or fewer products
Highlighted products that are offered include:
StemPro® MSC SFM: First serum-free medium (SFM) for growth and
expansion of human MSCs
MSC Differentiation Kits: Adipogenesis & Osteogenesis Differentiation Kits
for human MSCs
MesenPRO RS™ Medium: Reduced serum (2%) medium for mesenchymal
stem cell expansion
StemPro® Human Adipose - Derived Stem Cell Kit: Adult stem cells derived
from adipose tissue
Similar to BD Bioscience, Life Technologies offers links to 464 citation and reference
documents pertaining to mesenchymal stem cells, indicating that the company is
also positioning itself as a resource for mesenchymal stem cell relevant information.
F. LGC Standards/ATCC (www.lgcstandards-atcc.org) LCG Standards/ATCC offers nine products for culture of adult mesenchymal
stem cells, of which two are MSC primary cell populations.134 A summary of the
products is presented below.
Two are Primary Cells
“Adipose-Derived Mesenchymal Stem Cells; Normal, Human”
“Umbilical Vein-Derived Mesenchymal Stem Cells; Normal, Human”
One is “Mesenchymal Stem Cell Basal Medium”
One is a “Mesenchymal Stem Cell Growth Kit”
Five are research reagents
Dulbecco's Phosphate Buffered Saline (D-PBS)
Trypsin-EDTA for Primary Cells
134 LGC Standards/ATCC, Corporate Website. Available at: www. lgcstandards-atcc.org [Accessed May 9, 2013].
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Trypsin Neutralizing Solution
Gentamicin–Amphotericin B Solution
Penicillin–Streptomycin–Amphotericin B Solution Phenol Red
G. Lonza (www.Lonza.com)
Lonza began offering two mesenchymal stem cell products in 2009. In 2010, the
company grew its MSC product offerings to include 13 products that it designated as
MSC-specific, of which two were primary cell products.135 While other companies
have continued to expand their mesenchymal stem cell product lines, Lonza has not
added any new mesenchymal stem cell products since 2012.136 The company’s
selection of MSC products are presented below:
Poietics® Rat Mesenchymal Stem Cells
hMSC Human Mesenchymal Stem Cells
Human Adipose-Derived Stem Cells
hMSC Mesenchymal Stem Cell Chondrocyte Differentiation Medium
TheraPEAK™ MSCGM-CD™ Mesenchymal Stem Cell Medium, Chemically
Defined
TheraPEAK™ MSCGM-CD™ Mesenchymal Stem Cell Medium, Chemically
Defined
hMSC Mesenchymal Stem Cell Adipogenic Differentiation Medium
hMSC Mesenchymal Stem Cell Osteogenic Differentiation Medium
MSCGM™ Mesenchymal Stem Cell Growth Medium
Rat Mesenchymal Stem Cell Growth Medium Bullet Kit®
Rat MSC Adipogenic Differentiation Medium BulletKit®
Rat MSC Osteogenic Differentiation Medium BulletKit®
Nucleofector™ Kits for Human Mesenchymal Stem Cells (hMSC)
135 Lonza, Corporate Website. Available at: www.lonza.com [Accessed May 10, 2013]. 136 Ibid.
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H. Millipore (www.millipore.com)
Millipore currents markets 110 products under its “Mesenchymal Stem Cell” product
heading, for which the breakdown by category is shown below.137
MSC Products:
Antibodies (44 products)
Cell Culture Equipment & Supplies (42 products)
Multiwell Plates (12 products)
Kits & Assays (7 products)
Proteins & Enzymes (6 products)
Cell Culture Media & Reagents (6 products)
Cell Lines (2 products)
The two cells lines offered by the company are “Cryopreserved Rat Mesenchymal
Stem Cells (SCR027)” and “LT2 Immortalized Pancreatic Mesenchymal Cell Line
(SCR013).”
Millipore also offers 153 results in its Technical Library pertaining to mesenchymal
stem cells and 18 resources in its Learning Center.
I. Miltenyi Biotec (www. miltenyibiotec.com)
Miltenyi Biotec is a German bioscience company that pioneered MACS magnetic cell
separation technology in 1990, and later grew into a multinational team of more
than 1200 biomedical scientists, physicians, and engineers.138 As of 2010, the
company marketed 168 products as MSC-specific products. Now in 2012, the
company offers 199 products for MSC research, predominantly MSC-specific
antibodies, but also MSC Phenotyping Kits and MSC-qualified cell culture
137 Millipore, Corporate Website. Available at: www.millipore.com [Accessed May 11, 2013]. 138 Miltenyi Biotec. Corporate Website. “Company” Page: http://www.miltenyibiotec.com/en/NN_404_Company.aspx [Accessed May 10, 2013].
53
reagents.139 Clearly, the company is continuing to prioritize expansion of its
mesenchymal stem cell product line. The company has distributors that facilitate the
availability of its products on most continents.
J. PAA – The Cell Culture Company (www.paa.com)
PAA only markets five items as mesenchymal stem cell specific products, as shown in
the table below: Product Name, Catalog Number and Application.140
K. Promocell (www.promocell.com)
Promocell only markets 18 items as MSC-specific products, as shown in the chart on
the next page that presents: Product Name, Catalog Number, and Item
Description.141
139 Miltenyi Biotec, Corporate Website. Available at: www.miltenyibiotec.com [Accessed May 8, 2013]. 140 PAA – The Cell Culture Company, Corporate Website. Available at: www.paa.com [Accessed May 9, 2013]. 141 Promocell. Corporate Website. Available at: www.promocell.com [Accessed May 8, 2013].
PRODUCT NAME CATALOG NUMBER APPLICATION
FBS pre-tested on ES Cells A15-208 A supplement for growth of mesenchymal stem cells
MesenchymStem Medium U15-828 Complete medium for expansion of mesenchymal stem cells
DMEM, low Glucose, without L-glutamine E15-005
Base medium for growth of mesenchymal stem cells (glutamine must be added)
Accutase L11-007 For gentle dissociation of mesenchymal stem cells
Trypsin EDTA L11-004 For dissociation of mesenchymal stem cells
54
PRODUCT NAME CATALOG NUMBER ITEM DESCRIPTION
Human Mesenchymal Stem Cells from Adipose Tissue (hMSC-AT)
C-12977 500,000 cryopreserved cells
Human Mesenchymal Stem Cells from Bone Marrow (hMSC-BM)
C-12974 500,000 cryopreserved cells
Human Mesenchymal Stem Cells from Umbilical Cord Matrix (hMSC-UC)
C-12971 500,000 cryopreserved cells
Human Villous Mesenchymal Fibroblasts (HVMF)
C-12390 500,000 cryopreserved cells
Mesenchymal Stem Cell Growth Medium (Ready-to-use)
C-28010 500 ml
Human Mesenchymal Stem Cells from Adipose Tissue (hMSC-AT)
C-12978 500,000 proliferating cells
Human Mesenchymal Stem Cells from Bone Marrow (hMSC-BM)
C-12975 500,000 proliferating cells
Human Mesenchymal Stem Cells from Umbilical Cord Matrix (hMSC-UC)
C-12972 500,000 proliferating cells
Human Villous Mesenchymal Fibroblasts (HVMF)
C-12391 500,000 proliferating cells
Mesenchymal Stem Cell Adipogenic Differentiation Medium (Ready-to-use)
C-28011 100 ml
Mesenchymal Stem Cell Chondrogenic Differentiation Medium (Ready-to-use)
C-28012 100 ml
Mesenchymal Stem Cell Chondrogenic Differentiation Medium w/o Inducers (Ready-to-use)
C-28014 100 ml
Mesenchymal Stem Cell Osteogenic Differentiation Medium (Ready-to-use)
C-28013 100 ml
Mesenchymal Stem Cell Neurogenic Differentiation Medium (Ready-to-use)
C-28015 100 ml
hMSC-AT Pellet C-14092 > 1 million cells per pellet
hMSC-BM Pellet C-14090 > 1 million cells per pellet
hMSC-UC Pellet C-14091 > 1 million cells per pellet
HVMF Pellet C-14034 > 1 million cells per pellet
L. SA Biosciences (www.sabiosciences.com)
SA Biosciences, a Qiagen Company, is located in Frederick, Maryland, USA. The
company offers nine items that it categorizes as MSC-specific products, of which
only one appears to be truly specific for MSC research, their “Nano PreAMP cDNA
55
Synthesis Primer Mix - Human Mesenchymal Stem Cells. 142 This product is designed
for pre-amplification of cDNA from nanogram amounts of RNA (1-100ng) for multi-
gene expression analysis with RT² Profiler Human Mesenchymal Stem Cells PCR
Arrays.”143
A complete list of the products that the company designates as designed for MSC
research is shown below:
PRODUCT NAME
Nano PreAMP cDNA Synthesis Primer Mix - Human Mesenchymal Stem Cells
RT² Profiler™ PCR Array
RT² miRNA qPCR Primer Assay
RT² miRNA PCR Array
RT² qPCR Primer Assay
SureSilencing™ shRNA
Methyl Profiler DNA Methylation qPCR Assay
Multi-Analyte Profiler ELISArray™ Kits
Single Analyte ELISArray™ Kits
M. ScienCell Research Laboratories (www.sciencellonline.com)
ScienCell Research Laboratories, located in Carlsbad, CA, offers a more extensive line
of MSC-specific products.144 The company markets 80 products as designated as
specific for MSC research, including the following product types:
Primary MSCs derived from a variety of tissue sources and species
MSC Differentiation Kits (Osteogenic and Adipogenic)
Transfection Kits
MSC-specific cDNA Products
MSC-specific Total RNA Products
MSC-specific Genomic DNA Products
Wide variety of MSC-compatible cell culture tools and reagents
142 SA Biosciences, Corporate Website. Available at: www.sabiosciences.com [Accessed May 7, 2013]. 143 Nano PreAMP cDNA Synthesis Primer Mix - Human Mesenchymal Stem Cells – SA Biosciences – Product page. Available at: http://www.sabiosciences.com/rt_pcr_product/HTML/PBH-4082.html [Accessed May 6, 2013]. 144 ScienCell Research Laboratories, Corporate Website. Available at: www.sciencellonline.com [Accessed May 10, 2013].
56
A complete list of ScienCell’s MSC products is below:
PRODUCT NAME
Human Villous Mesenchymal Fibroblasts [HVMF]
Human Mesenchymal Stem Cells-bone marrow [HMSC-bm]
Human Mesenchymal Stem Cells-adipose [HMSC-ad]
Human Mesenchymal Stem Cells-hepatic [HMSC-hp]
Human Umbilical Mesenchymal Stem Cells [HUMSC]
Human Pulmonary Mesenchymal Stem Cells [HPMSC]
Human Vertebral Mesenchymal Stem Cells [HVMSC]
Mouse Mesenchymal Stem Cells-bone marrow [MMSC-bm]
Mesenchymal Stem Cell Chondrogenic Differentiation [MCDM]
Rat Mesenchymal Stem Cells-bone marrow [RMSC]
Horse Mesenchymal Stem Cells-adipose [HMSC-ad]
Dog Mesenchymal Stem Cells-adipose [DMSC-ad]
Mesenchymal Stem Cell Medium [MSCM]
Mesenchymal Stem Cell Osteogenic Differentiation M [MODM-500]
Mesenchymal Stem Cell Osteogenic Differentiation M [MODM-250]
Mesenchymal Stem Cell Adipogenic Differentiation M [MADM-500]
Mesenchymal Stem Cell Adipogenic Differentiation M [MADM-250]
Mesenchymal Stem Cell Medium-serum free [MSCM-sf]
Mesenchymal Stem Cell Medium -Phenol Red Free [MSCM-prf]
Mesenchymal Stem Cell Growth Supplement [MSCGS]
Mesenchymal Stem Cell Growth Supplement-serum free [MSCGS-sf]
Mesenchymal Stem Cell Transfection Kit [MesenFecta]
Human Villous Mesenchymal Fibroblast cDNA [HVMF cDNA]
Human Amniotic Mesenchymal Stromal Cell cDNA [HAMSC cDNA]
Human Chorionic Mesenchymal Stromal Cell cDNA [HCMSC cDNA]
Human Mesenchymal Stem Cell-bone marrow cDNA [HMSC-bm cD]
Human Mesenchymal Stem Cell-adipose cDNA [HMSC-ad cD]
Human Mesenchymal Stem Cell-hepatic cDNA [HMSC-hp cD]
Human Umbilical Mesenchymal Stem Cell cDNA [HUMSC cDNA]
Human Pulmonary Mesenchymal Stem Cell cDNA [HPMSC cDNA]
Human Vertebral Mesenchymal Stem Cell cDNA [HVMSC cDNA]
Human Villous Mesenchymal Fibroblast total RNA [HVMF tRNA]
Human Amniotic Mesenchymal Stromal Cell total RNA [HAMSC tRNA]
Human Chorionic Mesenchymal Stromal Cell total RNA [HCMSC tRNA]
Human Mesenchymal Stem Cell-bone marrow total RNA [HMSC-bm tR]
Human Mesenchymal Stem Cell-adipose total RNA [HMSC-ad tR]
Human Mesenchymal Stem Cell-hepatic total RNA [HMSC-hp tR]
Human Umbilical Mesenchymal Stem Cell total RNA [HUMSC tRNA]
Human Pulmonary Mesenchymal Stem Cell total RNA [HPMSC tRNA]
Human Vertebral Mesenchymal Stem Cell total RNA [HVMSC tRNA]
Human Villous Mesenchymal Fibroblast miRNA [HVMF miRNA]
Human Amniotic Mesenchymal Stromal Cell miRNA [HAMSC miRN]
Human Chorionic Mesenchymal Stromal Cell miRNA [HCMSC miRN]
Human Mesenchymal Stem Cell-bone marrow miRNA [HMSC-bm mi]
Human Mesenchymal Stem Cell-adipose miRNA [HMSC-ad mi]
Human Mesenchymal Stem Cell-hepatic miRNA [HMSC-hp mi]
Human Umbilical Mesenchymal Stem Cell miRNA [HUMSC miRN]
Human Pulmonary Mesenchymal Stem Cell miRNA [HPMSC miRN]
Human Vertebral Mesenchymal Stem Cell miRNA [HVMSC miRN]
Human Villous Mesenchymal Fibroblast Lysate [HVMFL]
57
Human Amniotic Mesenchymal Stromal Cell Lysate [HAMSCL]
Human Chorionic Mesenchymal Stromal Cell Lysate [HCMSCL]
Human Mesenchymal Stem Cell-bone marrow Lysate [HMSC-bm L]
Human Mesenchymal Stem Cell-adipose Lysate [HMSC-ad L]
Human Mesenchymal Stem Cell-hepatic Lysate [HMSC-hp L]
Human Umbilical Mesenchymal Stem Cell Lysate [HUMSCL]
Human Pulmonary Mesenchymal Stem Cell Lysate [HPMSCL]
Human Vertebral Mesenchymal Stem Cell Lysate [HVMSCL]
Human Villous Mesenchymal Fibroblast Genomic DNA [HVMF gDNA]
Human Amniotic Mesenchymal Stromal Cell Genomic DN [HAMSC gDNA]
Human Chorionic Mesenchymal Stromal Cell Genomic D [HCMSC gDNA]
Human Mesenchymal Stem Cell-bone marrow Genomic DN [HMSC-bm gD]
Human Mesenchymal Stem Cell-adipose Genomic DNA [HMSC-ad gD]
Human Mesenchymal Stem Cell-hepatic Genomic DNA [HMSC-hp gD]
Human Umbilical Mesenchymal Stem Cell Genomic DNA [HUMSC gDNA]
Human Pulmonary Mesenchymal Stem Cell Genomic DNA [HPMSC gDNA]
Human Vertebral Mesenchymal Stem Cell Genomic DNA [HVMSC gDNA]
Mesenchymal Stem Cell Growth Supplement-animal com [MSCGS-acf]
Mesenchymal Stem Cell Medium-serum free [MSCM-acf]
Mesenchymal Stem Cell Chondrogenic Differentiation [MCDS]
Mesenchymal Stem Cell Adipogenic Differentiation S [MADS]
Mesenchymal Stem Cell Adipogenic Differentiation S [MADS]
Mesenchymal Stem Cell Osteogenic Differentiation S [MODS]
Mesenchymal Stem Cell Osteogenic Differentiation S [MODS]
Mesenchymal Stem Cell Chondrogenic Differentiation [MCDM]
Mesenchymal Stem Cell Chondrogenic Differentiation [MCDM]
Mesenchymal Stem Cell Chondrogenic Differentiation [MCDS]
Mesenchymal Stem Cell Chondrogenic Differentiation [MCDS]
N. STEMCELL Technologies (www.stemcell.com)
STEMCELL Technologies, based in Vancouver, Canada, is a privately-owned stem cell
company that specializes in the production of research products. The company grew
out of the Media Preparation Service of the Terry Fox Laboratory for
Hematology/Oncology Research at the British Columbia Cancer Agency and now
provides more than 1000 research products to more than 70 countries worldwide.145
As such, it is not surprising that the company offers 20 mesenchymal stem cell
specific research products, as well as a “Mesenchymal Stem Cell Training Course”
that it sells alongside its research products.146 A complete listing of the company’s
products is shown below:
145 Stem Cell Technologies, Corporate Website. “About Us” Page: http://www.stemcell.com/en/About-Us.aspx. [Accessed May 12, 2013]. 146 Stem Cell Technologies, Corporate Website. Available at: www.stemcell.com [Accessed May 7, 2013].
58
PRODUCT NAME DESCRIPTION CATALOG #
Mesenchymal Stem Cell Training Course
Training course for mesenchymal stem cells 209
RosetteSep® Human Mesenchymal Stem Cell Enrichment Cocktail
Immunodensity isolation of untouched mesenchymal stem cells
15128
Fetal Bovine Serum for Human Mesenchymal Stem Cells
Prescreened FBS for human mesenchymal stem cells
6472
Fetal Bovine Serum for Human Mesenchymal Stem Cells
Prescreened FBS for human mesenchymal stem cells
6471
RosetteSep® Human Mesenchymal Stem Cell Enrichment Cocktail
Immunodensity isolation of untouched mesenchymal stem cells
15168
EasySep® Mouse Mesenchymal Stem/Progenitor Cell Enrichment Kit
Immunomagnetic Isolation of untouched mouse mesenchymal stem/progenitor cells
19771
MesenCult® Stimulatory Supplements (Human)
Supplement for detection of CFU-F and expansion of human mesenchymal stem cells
5402
MesenCult® Stimulatory Supplements (Mouse)
Supplement for detection of CFU-F and expansion of mouse mesenchymal stem cells
5502
MesenCult®-ACF Dissociation Kit Animal component-free dissociation kit for human mesenchymal stem cells
5426
MesenCult®-XF Medium Defined, xeno-free medium for human mesenchymal stem cells
5420
Adipogenic Stimulatory Supplements (Mouse)
Supplement for differentiating mouse mesenchymal stem cells into adipocytes
5503
MesenCult® Adipogenic Stimulatory Supplements (Human)
Supplement for differentiating human mesenchymal stem cells into adipocytes
5403
MesenCult®-XF Attachment Substrate Xeno-free attachment substrate for human mesenchymal stem cells
5424
MesenCult® MSC Basal Medium (Mouse)
Basal medium for mouse mesenchymal stem cells
5501
MesenCult® Proliferation Kit (Human) Medium for detection of CFU-F and expansion of human mesenchymal stem cells
5411
MesenCult® Osteogenic Stimulatory Kit (Human)
Kit for differentiating human mesenchymal stem cells into osteogenic progenitors
5404
MesenCult® Proliferation Kit (Mouse) Medium for detection of CFU-F and expansion of mouse mesenchymal stem cells
5511
Osteogenic Stimulatory Kit (for MSCs cultured in MesenCult-XF)
Kit for differentiating human mesenchymal stem cells cultured in MesenCult®-XF into osteogenic progenitors
5434
MesenCult® MSC Basal Medium (Human)
Basal medium for human mesenchymal stem cells
5401
MesenCult®-XF Culture Kit Defined, xeno-free culture kit for human mesenchymal stem cells
5429
59
A point of interest about StemCell Technologies is that the company offers products
for the enrichment and culture of both human and mouse mesenchymal stem cells,
advertised under its MesenCult® brand name. These products contain prescreened
supplements and are specially formulated to support the growth of mesenchymal
stem cells.
O. Thermo Scientific (www.thermoscientific.com)
Thermo Scientific, based in Suwanee, GA, offers a range of products for the study of
mesenchymal stem cells products, advertised under its Hyclone™ brand name.147 At
this time, the company offers only products for human mesenchymal stem cell
research.
147 Thermo Scientific, Corporate Website: www.thermoscientific.com. Accessed May 7, 2013.
PRODUCT NAME DESCRIPTION
HyClone AdvanceSTEM Mesenchymal Stem Cell Expansion Kit
Formulated to support expansion and maintenance of undifferentiated hMSCs.
HyClone Human Mesenchymal Stem Cell Screened Fetal Bovine Serum, US Origin
Prescreened and ready to use
HyClone CET Human Wharton’s Jelly Mesenchymal Stem Cells
Expanded and isolated from the inner portion of the human umbilical cord.
HyClone CET Human Adipose-Derived Mesenchymal Stem Cells
Isolated and expanded from human lipoaspirate using enzymatic treatments.
HyClone* CET Human Bone Marrow Mesenchymal Stem Cells
Isolated from human red bone marrow collected during a bone marrow aspiration procedure.
HyClone CET Human Amniotic Mesenchymal Stem Cells
HyClone AdvanceSTEM Osteogenic Differentiation Kit
Specifically developed for MSC research.
HyClone AdvanceSTEM Human Somatic Stem Cell Media
Specifically developed for MSC research.
HyClone CET Human Cord Blood CD133+ Stem Cells
Unlike Mesenchymal Stem Cells, Thermo Scientific* HyClone CET Human Cord Blood CD133+ Stem Cells, HCBHSC are mainly used in animal models to study the immune system and cardiovascular or circulatory defects.
HyClone AdvanceSTEM Adipogenic Differentiation Kit
Developed for applications in mesenchymal stem cell research.
60
P. R&D Systems (www.rndsystems.com)
R&D Systems, based in Minneapolis, MN, offers 18 MSC-specific products, which include:148
148 R&D Systems, Corporate Website. Available at: www.rndsystems.com [Accessed May 12, 2013].
PRODUCT NAME
ANTIBODY CATALOG NUMBER
APPLICATION KEY
Human Mesenchymal Stem Cell Functional Identification Kit SC006
Mouse Mesenchymal Stem Cell Functional Identification Kit SC010
Rat Mesenchymal Stem Cell Functional Identification Kit SC020
Rat Mesenchymal Stem Cells (1 x 10e6 cells/vial) PSC003
Human/Mouse StemXVivo Chondrogenic Base Media CCM005
Human/Mouse StemXVivo Chondrogenic Supplement, 0.5 mL CCM006
Human StemXVivo Osteogenic Supplement, 12.5 mL CCM008
Mouse StemXVivo Osteogenic Supplement, 12.5 mL CCM009
StemXVivo Mesenchymal Stem Cell Expansion Media
For use with human, mouse, and rat CCM004
Human Multi-potent Mesenchymal Stromal Cell Marker Ab Panel
Contains 25 ug each of antibodies to Stro-1, CD90, CD106, CD105, CD146, CD166, CD44, CD19, and CD45.
SC017
Human Multi-potent Mesenchymal Stromal Cell 4-Color Flow Kit
Contains conjugated antibodies to CD105-PerCp (Clone 166707), CD146-Fluorescein (Clone 128018), CD90-APC (CloneThy-1A1), and CD45-PE (Clone ICRF 2D1)
FMC002
Mouse Multi-potent Mesenchymal Stromal Cell 4-Color Flow Kit
Contains conjugated antibodies to CD105-Fluorescein (Clone 209701), CD29-PE (Clone 265917), Sca-1-APC (Clone 177228), and CD45-PerCP (Clone 30-F11)
FMC003
Mouse Multi-potent Mesenchymal Stromal Cell Marker Ab Panel
Contains 25 ug each of antibodies to Sca-1, CD106, CD105, CD73, CD29, CD44, CD11b, and CD45
SC018
Human/Mouse StemXVivo Osteogenic/Adipogenic Base Media CCM007
Mouse Patched 1/PTCH Affinity Purified Polyclonal Ab, Goat IgG FC, WB AF4105
Mouse Patched 1/PTCH MAb (Clone 413213), Rat IgG2A WB MAB4105
Recombinant Mouse EGF-L6, CF 4329-EG-025
Mouse Patched 1/PTCH MAb (Clone 413220), Rat IgG2A IHC, WB MAB41051
61
VIII. COMPANIES OFFERING RODENT MSC RESEARCH PRODUCTS
As mentioned earlier, there are 16 companies that offer human mesenchymal stem cell
products and eight that offer rat mesenchymal stem cell products. Not surprisingly,
there is significant overlap between the two groups, with the majority of MSC research
product suppliers offering research tools for both species.
A complete listing of rat MSC product providers is presented below, in alphabetical
order:
Cell Applications (www.cellapplications.com)
Celprogen (www.celprogen.com)
Cyagen Biosciences (www.cyagen.com) – China only
Genlantis (www.genlatis.com)
Millipore (www.millipore.com)
Miltenyi Biotec (www.miltenyibiotec.com)
ScienCell Research Laboratories (www.sciencellonline.com)
Trevigen (www.trevigen.com)
However, of these companies, only three of them have limited product offerings that
consist solely of rat MSC products. These companies are: Cell Applications, Genlantis,
and Trevigen. As these companies were not profiled in the previous section, they are
profiled here.
A. Cell Applications (www.cellapplications.com)
Cell Applications, located in San Diego, CA, is a specialty cell culture company. Its
focus is on providing high purity, characterized primary cells, including over 80
different human and animal primary cell types, cell growth media and culture
reagents, and cell marker and signaling antibodies. The company began offering
mesenchymal stem cell products in 2008, and by 2010, it was offering six MSC-
relevant research products, shown on the following page:149
149 Cell Applications - Corporate Website. Available at: www.cellapplications.com [Accessed May 9, 2013].
62
In late 2011, the company added a seventh MSC product to its product offerings,
“Human Marrow Stromal Cell Media.”
For this company, it is important to note the use of alternative product
nomenclature, “Marrow Stromal Cell,” in place of the more commonly recognized
term “Mesenchymal Stem Cell.” It is likely that the company loses a significant
amount of online search traffic due to this languaging decision and also experiences
a reduction in general product interest from those that do reach the site. Also, the
company is somewhat unique in offering research products from a range of animal
species – specifically, canine and feline, in addition to rat.
B. Genlantis (www.genlantis.com)
Genlantis, located in San Diego, CA, is a research products company that specializes
in GenePORTER® brand transfection products for gene delivery and plasmid DNA, as
well as GeneSilencer® brand reagents useful in the delivery of siRNA for gene
suppression in mammalian cells.
However, even though gene delivery into mesenchymal stem cells represents an
area of intense therapeutic interest, Genlantis has not yet applied its branded
technologies to its mesenchymal stem cell product offerings. Rather, the company
currently offers a relatively straight-forward and limited selection of MSC products,
which it introduced in 2009.
PRODUCT NAME
Canine Marrow Stromal Cell Media
CnMSC: Canine Marrow Stromal Cells
Feline Marrow Stromal Cell Media
Feline Marrow Stromal Cells
Rat Marrow Stromal Cell Media
Rat Marrow Stromal Cells: RMSC
63
The five MSC products Genlantis currently offers are shown below:150
C. Trevigen, Inc. (www.trevigen.com) Trevigen, Inc., located in Gaithersburg, MD, also offers five MSC-specific products,
shown below:151
PRODUCT NAME CATALOG #
Rat Mesenchymal Cells 5000-001-01
Rat Mesenchymal Stem Cell Starter Kit 5000-001-K
Rat Mesenchymal Replenisher Kit 5000-001-R
Mesenchymal Stem Cell Adipogenic Differentiation Kit 5010-024-K
Mesenchymal Stem Cell Osteogenic Differentiation Kit 5011-024-K
Interestingly, Trevigen heavily pre-marketed its “Rat Mesenchymal Adipogenic” and
“Rat Osteogenic Differentiation” Kits in mid-2009, and launched them in November
of that year, at the same time that it also made available primary “Rat Mesenchymal
Stem Cells.” In late spring 2010, Trevigen then launched two more products, a “Rat
Mesenchymal Stem Cell Starter Kit” and a “Rat Mesenchymal Replenisher Kit.”
It will be interesting to see how the company continues its rat mesenchymal stem
cell product development in 2011, and to observe whether or not the company
decides to extend its reach into offering human mesenchymal stem cell products as
well.
While the company is a broad-based research products company, it has a preference
for product development in the form of kits, and this preference is demonstrated
here in its decision to launch five rat mesenchymal stem cell products, of which four
of those products are in kit format.
150 Genlantis - Corporate Website. Available at: www.genlantis.com [Accessed May 12, 2013]. 151 Trevigen, Inc., Corporate Website. Available at: www.trevigen.com [Accessed May 11, 2013].
PRODUCT NAME
Rat Mesenchymal Stem Cell (RMSC), Complete System
Rat Mesenchymal Stem Cell (RMSC), vial
Rat Mesenchymal Stem Cell (RMSC), Medium
Rat Osteoblast Differentiation Medium 250 ml
Rat Adipocyte Differentiation Medium 250 ml
64
IX. ANALYSIS OF MSC RESEARCH ACTIVITY
A. By Tissue Source of Origin
It is also relevant to consider the breakdown of mesenchymal stem cell (MSC)
research activity by tissue source, in order to evaluate researcher preferences for
MSCs derived from a variety of sources.
As mentioned previously, MSC are a well-characterized population of adult stem
cells, for which 12,547 scientific articles have been published to date.152 Of those,
approximately one-third (5,614) of the articles were published within the past two
years. Using this population of MSC-specific publications from the trailing 24 months
(January 1, 2010 through January 1, 2012), the following is a breakdown of MSC
research activity by tissue source of origin, with results presented in alphabetical
order.
152 Figure calculated using search of Pubmed Database (http://www.ncbi.nlm.nih.gov/pubmed) for the search terms: ["Mesenchymal Stem Cell" OR "Mesenchymal Stem Cells"] OR ["Marrow Stromal Cell" OR "Marrow Stromal Cells"] OR ["Multi-potent Stromal Cell" OR "Multi-potent Stromal Cells"] OR ["Colony-Forming Unit-fibroblast" OR "Colony-Forming Unit-fibroblasts"]. Search encompassed four technical terms for the cell type, as well as variations in singular versus plural usage of terminology. Executed March 10, 2012.
65
TABLE: Percent of Mesenchymal Stem Cell Research Activity by Tissue Source
ADULT SOURCE (ALPHABETICAL) % OF MSC RESEARCH BY TISSUE SOURCE
Adipose Tissue 22.1
Aortic Artery 0.1
Articular Cartilage 0.2
Bone Marrow 53.1
Dental Pulp 2.7
Kidney glomeruli 0.4
Liver 0.6
Lung 0.6
Neural Tissue 0.4
Pancreas 0.3
Periostium 0.6
Skeletal Muscle 1.4
Skin 2.7
Stroma of Spleen 1.1
Stroma of Thymus 1.3
Synovium and Synovial Fluid 0.1
Tendons <0.1
Trabecular Bone <0.1
Vena Cava <0.1
Xiphoid Cartilage <0.1
TOTAL OF ADULT SOURCES = 87.7
FETAL SOURCE (ALPHABETICAL) % OF MSC RESEARCH BY CELL TYPE
Amnion 2.1
Fetal Tissues (Pancreas, Spleen, Thymus) <0.1
Placenta 1.7
Umbilical Cord Vasculature 8.3
Wharton's Jelly 0.2
TOTAL OF FETAL SOURCES = 12.3
COMBINED TOTAL (ADULT AND FETAL) = 100
66
The first interesting finding from this data is the relative frequency of MSC data
performed with cells from adult versus fetal sources; notice that research with adult-
derived MSCs is nearly eight times as common as research with fetal-derived MSCs.
The second interesting finding is the high relative frequency of MSC research
performed with bone marrow-derived (53.1%) and adipose-derived (22.1%) MSCs.
Umbilical cord vasculature-derived MSCs also represent a substantial percentage of
MSC research (8.3%), although this value is substantially lower than rates of
research on bone-marrow derived and adipose-derived MSCs. After considering
those tissue sources of origin, research activity utilizing mesenchymal stem cells
from all other sources becomes relatively evenly distributed at or around 2% or less.
Below, the data is presented again, this time in descending order of frequency (as
opposed to alphabetically), with tissue sources contributing less than 1% of MSC
research activity combined into general category headings of “Adult–Other Sources”
and “Fetal-Other Sources.”
87.7%
12.3%
0.0%
10.0%
20.0%
30.0%
40.0%
50.0%
60.0%
70.0%
80.0%
90.0%
100.0%
Adult Sources Fetal Sources
Percent of MSC Research
67
ADULT SOURCE % OF MSC RESEARCH PERFORMED
WITH CELL TYPE
Bone Marrow 53.1
Adipose Tissue 22.1
Dental Pulp 2.7
Skin 2.7
Skeletal Muscle 1.4
Stroma of Thymus 1.3
Stroma of Spleen 1.1
Adult - Other Sources 3.3
TOTAL OF ADULT SOURCES = 87.7
FETAL SOURCE % OF MSC RESEARCH PERFORMED
WITH CELL TYPE
Umbilical Cord Vasculature 8.3
Amnion 2.1
Placenta 1.7
Fetal - Other Sources 0.2
TOTAL OF FETAL SOURCES = 12.3
COMBINED TOTAL (ADULT AND FETAL) = 100
Finally, the data is presented a last time, again in descending order of frequency, but
with adult and fetal sources combined into a single chart and graphically
represented.
SOURCE % OF MSC RESEARCH BY CELL TYPE
Bone Marrow 53.1
Adipose Tissue 22.1
Umbilical Cord Vasculature 8.3
Adult - Other Sources 3.3
Dental Pulp 2.7
Skin 2.7
Amnion 2.1
Placenta 1.7
Skeletal Muscle 1.4
Stroma of Thymus 1.3
Stroma of Spleen 1.1
Fetal - Other Sources 0.2
TOTAL = 100
68
B. By Clinical Application
Next, consider a breakdown of MSC research activity by clinical application, also
produced using the described trailing 24-month data set:
TABLE: Percent of Mesenchymal Stem Cell Research by Clinical Application
CLINICAL APPLICATION % OF MSC RESEARCH
Bone Tissue Engineering 30.1
Cartilage Repair 17.5
Wound Healing 17.0
Cardiac Repair 13.1
Muscle Repair 6.1
Tendon Repair 2.9
Neural Repair 2.7
Osteoarthritis Therapy 2.6
Eye (Corneal) Repair 0.1
Other 7.9
TOTAL 100%
0.2%1.1%
1.3%1.4%
1.7%2.1%
2.7%2.7%
3.3%
8.3%
22.1%
53.1%
MSC Research by Cell Type
Fetal - Other Sources
Stroma of Spleen
Stroma of Thymus
Skeletal Muscle
Placenta
Amnion
Skin
Dental Pulp
Adult - Other Sources
Umbilical Cord Vasculature
Adipose Tissue
Bone Marrow
69
C. By Species Source
Next, consider a breakdown of mesenchymal stem cell research activity by species
source, again produced using the described trailing 24 month data set:
TABLE: Percent of Mesenchymal Stem Cell Research by Species Source
SPECIES SOURCE % OF MSC RESEARCH
Human 40.1
Rat 28.7
Mouse 28.5
Pig 0.9
Sheep 0.3
Canine 0.2
Goat 0.2
Other 1.1
TOTAL = 100.0%
0.1% 2.6% 2.7% 2.9%
6.1%
7.9%
13.1%
17.0%17.5%
30.1%
MSC Research by Clinical Application
Eye (Corneal) Repair
Osteoarthritis Therapy
Neural Repair
Tendon Repair
Muscle Repair
Other
Cardiac Repair
Wound Healing
Cartilage Repair
Bone Tissue Engineering
70
0.2% 0.2% 0.3% 0.9% 1.1%
28.5%
28.7%
40.1%
MSC Research by Species Source
Canine
Goat
Sheep
Pig
Other
Mouse
D. By Geographical Region
Finally, mesenchymal stem cell publications from the trailing 24 months are
presented below, with mesenchymal stem cell research activity assessed by region.
TABLE: Percent of Mesenchymal Stem Cell Research Activity By Geographical Region
REGION % OF MSC RESEARCH
USA 30.8
Europe 28.7
Asia 22.2
Australia/NZ 4.3
Canada 3.9
India 3.4
South America 2.1
Mexico 0.5
Other 4.1
TOTAL = 100.0%
The intriguing finding from this data is the concentrated study of mesenchymal stem
cells in the United States, Europe, and Asia.
71
30.8%
28.7%
22.2%
4.3%3.9%
4.1%3.4% 2.1% 0.5%
MSC Research by Geographical Region
USA
Europe
Asia
Australia/NZ
Canada
Other
India
South America
Mexico
72
X. MARKET TREND ANALYSIS
A. Scientific Publication Rate Analysis
1. Historical 10-Year Analysis
PubMed is a service of the US National Library of Medicine that is a meta-
database containing citations from MEDLINE and a diverse range of other life
science journals.153 This section evaluates rate data over a historical ten year
period (“Trailing 10 Year Analysis”), using PubMed for aggregated analysis.
Interestingly, publication rate analyses show a relatively steady, linear
progression from 2001 through 2008, and then a sharp, spiked increase during
the years of 2009 through 2011. Several reasons may exist for this, as
summarized below:
Concern over the availability of funding for human embryonic stem cell
(hESC) research has continued to grow over the past two years, made
worse by the August 2010 federal court ruling against embryonic stem
cell research, which overturned President Obama's 2009 executive order
that had eased limits on such funding. These funding concerns may be
prompting researchers to transition into areas of adult stem cell research,
including MSCs.
MSCs are characterized by relative ease of isolation, as well as
advantages in terms of availability, expandability, and transplantability.
Characteristics that make a cell type convenient for research use will
keep existing researchers involved with the cell type, as well as attract
new interest.
153 PubMed Service - US National Library of Medicine. Available at: www.pubmed.com [Accessed March 15, 2012].
73
Immuno-advantaged characteristics of MSCs allow in vitro research
advances to be smoothly transitioned into pre-clinical and clinical
research studies. As such, examples such as the multi-million dollar
funding Mesoblast is receiving from Cephalon, Inc., in the form of a
strategic alliance, is an example of large-scale research and development
(R&D) dollars being invested into the cell type.
The time period from 2008 to 2011 represented a period during which
the number of research supply companies offering MSC products grew
significantly. When MSC research products emerged in 2006, only three
commercial suppliers of MSC products were available. As of 2008, eight
companies were offering research products specific for MSC study. Now,
there are 16 companies that offer human MSC products and eight that
offer rat MSC products. Taking into account the significant overlap in
providers of human and rat MSC products, there are currently 19
commercial suppliers offering specialty research tools to facilitate the
study of MSCs. In addition, nearly all antibody specialty companies now
offer MSC-specific antibodies. Of these, Abcam has developed a
comprehensive mesenchymal stem cell product line composed of specific
antibodies (40 products), proteins and peptides (four products), and kits
(two products) specialized for mesenchymal stem cell research, and the
company markets these products under a designated “Mesenchymal
Stem Cell” Product Category on its website.154
154 Abcam, Corporate Website. “Mesenchymal Stem Cell” Product Listings. Available at: http://www.abcam.com/index.html?pageconfig=productmap&cl=3427 [Accessed December 1, 2010].
74
# of Mesenchymal Stem Cell Publications, by Year
0
2000
4000
6000
8000
10000
12000
14000
2000 2002 2004 2006 2008 2010 2012
Year
# o
f M
SC
Pu
blicati
on
s
TABLE: PUBMED Analysis of MSC Publication Rates by Year
YEAR # of PUBLICATIONS
2002 139
2003 292
2004 519
2005 810
2006 1,085
2007 1,436
2008 1,738
2009 2,746
2010 5,821
2011 12,547
In 2010, the annual growth rate for MSC research publications was 112%.
In 2011, the annual growth rate for MSC research publications was 116%.
75
2. Future 5-Year Projection
Using an exponential best-fit line (which is more accurate than a linear,
polynomial, logarithmic, power, or moving average best-fit line), with an r2 value
of 0.9899155, five-year projection data predicts that mesenchymal stem cell
publications will reach approximately 73,308 annually by 2016. If the exponential
growth observed over the previous five years continues, the predicted increase
in publication rates for the coming five years would represent approximately a
5.8-fold increase in published mesenchymal stem cell research activity from
2011 to 2016.
TABLE: PUBMED Analysis of Mesenchymal Stem Cell Publication Rates by Year, with 5-Year Projections
YEAR # of PUBLICATIONS
2002 139
2003 292
2004 519
2005 810
2006 1,085
2007 1,436
2008 1,738
2009 2,746
2010 5,821
2011 12,547
2012 20,021
2013 30,009
2014 42,289
2015 58,223
2016 73,308
*Note: Numbers in italics are projections.
155 r2 is a statistical term that represents how good one term is at predicting another, in this case, how well the year can predict # of publications using the best-fit line set to the trailing five-year data. If r2 is 1.0 then given the value of one term, you can perfectly predict the value of the other term. If r2 is 0.0, then knowing one term does not help you predict the other term at all.
76
# of Mesenchymal Stem Cell Publications, by Year
y = 1065.4x2 - 4E+06x + 4E+09
R2 = 0.9899
0
10000
20000
30000
40000
50000
60000
70000
80000
2007 2009 2011 2013 2015 2017
Year
# o
f M
SC
Pu
blicati
on
s
PUBMED Analysis of MSC Publication Rates by Year,
Best Fit Line Equation:
y = 1065.4x2 – 4E+06x + 4E+09 R2 = 0.9899
77
B. MSC Grant Rate Analysis
1. Historical 10-Year Analysis
CRISP (Computer Retrieval of Information on Scientific Projects)156 is a
searchable database of federally funded biomedical research projects conducted
at universities, hospitals, and other research institutions. The database,
maintained by the Office of Extramural Research at the National Institutes of
Health (NIH), includes projects funded by the NIH, Substance Abuse and Mental
Health Services (SAMHSA), Health Resources and Services Administration (HRSA),
Food and Drug Administration (FDA), Centers for Disease Control and Prevention
(CDCP), Agency for Health Care Research and Quality (AHRQ), and Office of the
Assistant Secretary of Health (ASH). Users can use the CRISP interface to search
for grants funding specific types of scientific research, such as “Mesenchymal
Stem Cell” 157 research, which is shown below.
TABLE: CRISP Analysis of Grant Rates per Year (Trailing 10-Year Analysis)
YEAR # OF GRANTS
2002 45
2003 83
2004 92
2005 97
2006 117
2007 105
2008 117
2009 154
2010 194
2011 229
156 Computer Retrieval of Information on Scientific Projects (CRISP). Available at: http://crisp.cit.nih.gov/ [Accessed December 1, 2012]. 157 Again, the following set of comprehensive search terms was used: ["Mesenchymal Stem Cell" OR "Mesenchymal Stem Cells"] OR ["Marrow Stromal Cell" OR "Marrow Stromal Cells"] OR ["Multi-potent Stromal Cell" OR "Multi-potent Stromal Cells"] OR ["Colony-Forming Unit-fibroblast" OR "Colony-Forming Unit-fibroblasts"]. Executed December 1, 2012.
78
CRISP Analysis of Mesenchymal Stem Cell Grant Rates per Year
0
50
100
150
200
250
2002 2007 2012
Year
# o
f G
ran
ts
From 2010 to 2011, the annual growth rate for mesenchymal stem cell grants
increased by 18%.
2. Future 5-Year Projection Below is five-year projection data for grant rates. Notice near doubling is
predicted to occur within a five year time period (229 publications in 2011 to 417
publications in 2016).
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TABLE: MSC Grant Rate Analysis (Future Projection)
YEAR # OF GRANTS
2002 45
2003 83
2004 92
2005 97
2006 117
2007 105
2008 117
2009 154
2010 194
2011 229
2012 251
2013 289
2014 329
2015 372
2016 417
*Note: Numbers in italics are projections.
y = 1.5985x2 - 6397.8x + 6E+06R² = 0.9258
0
50
100
150
200
250
300
350
400
450
2000 2005 2010 2015 2020
# o
f G
ran
ts
Year
CRISP Analysis of Mesenchymal Stem Cell Grant Rates per Year
80
Since the best-fit line for MSC grants is exponential (not linear, polynomial,
logarithmic, power, or a moving average best-fit line), with an r2 value of 0.9258,
the annual growth rate is expected to increase each year over the coming five
year period. However, it should be noted that r2 value for this best-fit line is only
0.9258, which means that even an appropriately fitted trend line is not entirely
descriptive of the data set. In the graph above, the observed data points do
show substantial variation around the trend line.
While a 18% year-over-year increase seems subdued in comparison to the year-
over-year rate of increase observed for scientific publications (116% from 2010
to 2011), a compounded annual growth rate of 18% is a powerful force. Also, it
should be considered that grant rates are highly dependent on external factors
(economic stability and available government funding), while scientific
publication rates are free of these constraining factors. For this reason,
publication rate analysis is generally considered a “gold standard” approach for
accurate prediction of research activity and trends within scientific communities.
CRISP Analysis of Grant Rates per Year
y = 1.5985x2 - 6397.8x + 6E+06 R2 = 0.9258
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C. Patent Analysis
Similar to the CRISP system utilized in the previous section for analysis of
mesenchymal stem cell grant data, the United States federal government supports a
searchable patent database, the “United State Patent and Trademark Office (USPTO)
Full-Text and Image Database.”158
For purposes of keeping this analysis specific to mesenchymal stem cells, the
following terms were screened for their presence within the Title or Abstract of U.S.
patents: ["Mesenchymal Stem Cell" OR "Mesenchymal Stem Cells"]. The Title and
Abstract were chosen as the areas of the patent to search, because the Title is a
highly-specific phrase that captures the focus of the patent and the Abstract is
limited to a 2-3 sentence description, making it highly-focused and specific as well.
As of 2013, USPTO patent search determines that there are 69 patents that have the
above terms appearing within the Title or the Abstract of the patent.159 A complete
list of these patents is shown on the next page.
For reference, where the MSC search terms above are searched within all fields of all
U.S. patents issued to date (including within the Description, Specifications, Claims
and more), 541 total results are returned. However, these patents are not included
in the list below, because in these situations MSCs are often referenced within a
patent for purposes of describing prior methods, knowledge, or systems.
158 United States USPTO Patent Full-Text and Image Database. Available at: http://www.uspto.gov/patents/process/search/#heading-1. [Accessed June 25, 2013.] 159 United States USPTO Patent Full-Text and Image Database (PatFT): http://www.uspto.gov/patents/process/search/#heading-1. The following set of comprehensive search terms was used: ["Mesenchymal Stem Cell" OR "Mesenchymal Stem Cells”]. Executed June 25, 2013.
82
TABLE. Mesenchymal Stem Cell Specific U.S. Patents (Search Terms Appearing within Title or Abstract of Patent; USPTO Full-Text and Image Database” Search)
PATENT # TITLE CATEGORY
8,465,733 Pharmaceutical composition containing human mesenchymal stem cell
Drug Discovery & Development
8,455,254 Method of accelerating osteogenic differentiation and composition thereof Differentiation
8,455,245 ABCB5 positive mesenchymal stem cells as immunomodulators Other
8,450,271 Peptide-based scaffolds for cartilage regeneration and methods for their use Other
8,445,279
Cultured cell construction containing spheroids of mesenchymal stem cells and utilization thereof
Other
8,444,968 Cartilage repair methods Cellular Therapy
8,440,199 Methods for mobilizing mesenchymal stem cells in a patient Cellular Therapy
8,440,177
Method of treating graft versus host disease using adipose derived mesenchymal stem cells
Cellular Therapy
8,435,509
Creation of a biological atrioventricular bypass to compensate for atrioventricular block
Cellular Therapy
8,431,400 Dermal sheath cup cell population Other
8,431,397
Differentiation of human mesenchymal stem cells to cardiac progenitor cells that promote cardiac repair
Differentiation
8,388,947 Maintenance and propagation of mesenchymal stem cells Cell Culture
8,361,791
Mesenchymal stem cells with increased developmental potency by expressing Nanog
Differentiation
8,361,485 Conditioned cell culture medium compositions and methods of use Cell Culture
8,343,922
Compositions and methods for the stimulation or enhancement of bone formation and the self-renewal of cells
Differentiation
8,318,495 Hematopoietic progenitor cell gene transduction Other
8,309,095
Parathyroid hormone receptor activation and stem and progenitor cell expansion
Cell Expansion
8,287,853 Methods of culturing mesenchymal stem cells Cell Culture
8,278,101 Stem cell bioprocessing and cell expansion Cell Expansion
8,277,795
Methods and compositions for treating motor neuron diseases comprising mesenchymal stem cells
Cellular Therapy
8,222,216 Mesenchymal cell proliferation promoter and skeletal system biomaterial Differentiation
8,216,839
Systems and methods for making hepatocytes from extrahepatic somatic stem cells and use thereof
Differentiation
8,202,551
Tissue engineered cartilage, method of making same, therapeutic and cosmetic surgical applications using same
Differentiation
8,192,988 Methods for increasing potency of adult mesenchymal stem cells Cell Expansion
8,178,084 Pharmaceutical kits comprising mesenchymal stem cells
Drug Discovery & Development
8,163,495 Method for isolating and/or identifying mesenchymal stem cells (MSC) Cell Culture
8,158,121 Cardiac muscle regeneration using mesenchymal stem cells Cellular Therapy
8,147,803 Ultrasmall superparamagnetic iron oxide nanoparticles and uses thereof Other
8,142,774 Methods of treatment using electromagnetic field stimulated stem cells Cellular Therapy
8,142,773
Methods of implanting mesenchymal stem cells for tissue repair and formation
Cellular Therapy
8,138,147 Conditioned cell culture medium compositions and methods of use Cell Culture
8,137,688
Hydroxyphenyl cross-linked macromolecular network and applications thereof
Other
8,119,397 Therapeutic agents and therapeutic methods for treating injured tissue Cellular Therapy
83
8,110,400
Culture of mammalian pluripotent stem cells in the presence of hyaluronan induces differentiation into multi-lineage progenitor cells
Cell Culture
8,105,832 Methods for modifying stem cell characteristics Differentiation
8,105,791 Marker for stem cells and its use Cell Culture
8,084,023 Maintenance and propagation of mesenchymal stem cells Cell Expansion
8,039,256 Culturing method of mesenchymal stem cell Cell Culture
8,017,390
Method for the preparation of dermal papilla tissue employing mesenchymal stem cells
Differentiation
7,968,126
Creation of a biological atrioventricular bypass to compensate for atrioventricular block
Cellular Therapy
7,943,579
Osteogenic implant matrices and endosseous tooth implants with improved osteointegration properties
Cellular Therapy
7,943,136
Parathyroid hormone receptor activation and stem and progenitor cell expansion
Cell Expansion
7,915,039 Method for obtaining mesenchymal stem cells Cell Culture
7,897,727 Bioactive peptide for cell adhesion Other
7,892,829 Cardiac muscle regeneration using mesenchymal stem cells Cellular Therapy
7,879,603
Method for differentiating mesenchymal stem cells into steroid-producing cells
Differentiation
7,832,566
Method and apparatus for separating and concentrating a component from a multi-component material including macroparticles
Other
7,829,335 Method of differentiation induction to osteoblasts Differentiation
7,829,296
Kinesin polypeptides, polynucleotides encoding same and compositions and methods of using same
Other
7,807,462 Method for producing a functional neuron Differentiation
7,794,702
Mesenchymal stem cells as a vehicle for ion channel transfer in syncytial structures
Other
7,749,710 Marker for stem cells and its use Cell Culture
7,744,869
Methods of treatment using electromagnetic field stimulated mesenchymal stem cells
Cellular Therapy
7,741,114 Antibodies for identifying and/or isolating at least one cell population Cell Culture
7,732,203 Method for transdifferentiating mesenchymal stem cells into neuronal cells Differentiation
7,718,376 Identification and isolation of somatic stem cells and uses thereof Cell Culture
7,704,739
Method of isolating and culturing mesenchymal stem cell derived from umbilical cord blood
Cell Culture
7,678,884 Biologically active peptide and agent containing the same Other
7,635,477
Parathyroid hormone receptor activation and stem and progenitor cell expansion
Cell Expansion
7,615,374
Generation of clonal mesenchymal progenitors and mesenchymal stem cell lines under serum-free conditions
Cell Culture
7,592,176 Method of forming mesenchymal stem cells from embryonic stem cells Differentiation
7,592,174 Isolation of mesenchymal stem cells Cell Culture
7,582,477
Method of isolating and culturing mesenchymal stem cell derived from cryopreserved umbilical cord blood
Cell Culture
7,541,030
Antibodies for identifying and/or isolating at least one cell population which is selected from the group comprising haematopoietic stem cells, neuronal stem cells, neuronal precursor cells, mesenchymal stem cells and mesenchymal precursor cells
Cell Culture
7,534,607 Method of producing cardiomyocytes from mesenchymal stem cells Differentiation
7,514,074 Cardiac muscle regeneration using mesenchymal stem cells Cellular Therapy
7,504,099 Methods of inducing or enhancing connective tissue repair Cellular Therapy
7,476,540 Monoclonal antibodies to mesenchymal stem cells Cell Culture
7,371,818 Polypeptide and DNA thereof Other
84
The table below presents a categorical breakdown of U.S. patents with MSC terms
(“Mesenchymal Stem Cell” OR “Mesenchymal Stem Cells”) contained within the
Title or Abstract. It identifies that the most common type of MSC patents are those
pertaining to “Cell Culture” topics, followed by MSC patents about “Cell Expansion.”
See the table and chart below for additional detail.
TABLE. Categorical Breakdown of U.S. Patents with Mesenchymal Stem Cell Terms within the Title or Abstract
PATENT CATEGORY
# OF PATENTS WITH MSC TERMS IN TITLE OR
ABSTRACT % OF PATENTS
Cell Culture 18 26.1%
Cellular Therapy 15 21.7%
Differentiation 15 21.7%
Cell Expansion 6 8.7%
Drug Discovery & Development 2 2.9%
Other 13 18.8%
TOTAL 69 100.0%
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XI. MESENCHYMAL STEM CELLS AS TOOLS FOR DRUG DISCOVERY
The prospect of using mesenchymal embryonic stem cells for cell and gene therapy
applications has created significant public enthusiasm, but an attractive alternative in
the near future is the use of mesenchymal stem cells as tools for drug discovery.
In general, the market for stem cell assays to aid in early stage drug development is vast,
as stem cells differentiated into primary cells could form the basis of predictive assays
capable of filtering out compounds at an earlier stage of drug discovery. Historically,
the drug discovery and development process has been an extremely expensive and
time-consuming endeavor, requiring an investment of 12 to 15 years and a cost of over
$800 million per successful drug.160 Because unexpected side effects, usually involving
either the liver or heart, causes one in three drugs to fail toxicity requirements in clinical
trials, several companies are now focused on differentiating stem cells into hepatocytes
and cardiomyocytes for early-stage toxicity testing. To date, mesenchymal stem cells
have shown evidence that they may have the capability to serve as suitable models for
cardiotoxicity, hepatotoxicity, genotoxicity/epigenetic, and reproductive toxicology
screening.161
However, it is important to note that the main benefit of using stem cells for DNA
damage and toxicity testing is that current animal-based methods used to predict
toxicity are poorly predictive of human response. Thus, using human stem cells in
bioactivity and drug toxicity assays could dramatically reduce a key obstacle of
conventional drug development - that drug response differences between humans and
animals can be significant. Thus, when considering toxicology screening applications for
mesenchymal stem cells, it is important to consider that many researchers will have a
preference for working with human mesenchymal stem cell assays.
Nonetheless, there are several scenarios in which it would be commercially preferable
to develop mouse MSC screening assays in place of, or in conjunction with, human MSC
160 Technology & Discovery – Vistagen, Inc. – Corporate Website. Available at: http://www.vistagen.com/htm _pages/toxicity_screen.htm [Accessed December 1, 2010]. 161 Stem Cells in Research. Available at: www.nwabr.org/education/pdfs/StemCell/Inman_Stem_Cells_05.ppt [Accessed March 9, 2012].
86
screening assays. These situations are discussed in Section XI.B., “Potential Drug
Screening Advantages of Mouse MSCs.”
A. Companies Developing Toxicology Screening Platforms for Stem Cells
At this time, several companies are developing toxicology screening platforms for
stem cells. However, to date, none of them are using mesenchymal stem cells in
their applications. In the future, they may decide to do so. Companies involved with
this type of applied stem cell research include:
Advanced Stem Therapeutics: Generates stem cell lines for use in toxicology
screening, as well as disease research.
VistaGen, Inc.: Develops embryonic stem cell screening technologies for
drug discovery and toxicology assessment. VistaGen was awarded a Phase
1 Small Business Innovation Research (SBIR) research grant of $249,000 by
the NIH entitled “Development of Stem Cell Technologies for Toxicology
Assessment.”
BioE, Inc.: Sells a cord blood stem cell line called the Multi-Lineage
Progenitor Cell™ (MLPC™), of which the company announced the discovery
in 2004 and launched in July 2005.
Cellartis AB: Has more than 30 stem cell lines available for the discovery of
new pharmaceuticals, regenerative medicines, and toxicology. Two of the cell
lines are listed in the National Institutes of Health (NIH) Human Embryonic
Stem Cell Registry (www.nih.gov), and 20 are listed in the UK Stem Cell Bank
(http://www.ukstemcellbank .co.uk/).
Cellomics: Offers technology for using stem cells in single-cell, high-content
screening assays involving imaging and confocal microscopy.
87
Evotec Technologies: Also offers technology for using stem cells in high-
content screening assays involving imaging and microscopy.
B. Potential Drug Screening Advantages of Mouse MSCs
1. Acquisition, Handling, and Differentiation Advantages of Mouse MSCs
One way in which mouse MSCs could potentially be advantageous over human
MSCs for use in bioactivity and toxicity screening would be if they were viewed
by the scientific community as easier to acquire, handle, or differentiate.
For instance, there is evidence that the modulating factors required to
differentiate rat mesenchymal stem cells into their distinct mesenchymal
lineages are better defined for rat MSCs cells than for human MSCs, at least for
some lineages of interest.162 In one example of this principal, a group of
researchers at the Institute for Stem Cell Research in the U.K. chose to use
mouse stem cell-derived neural cells during development of a high-throughput
assay to screen for Alzheimer's disease.163
Additionally, because mesenchymal stem cells are derived from bone marrow of
a host species, it is usually easier to acquire rat MSC samples than human MSC
samples. Finally, there are less stringent patent restrictions for rat MSCs as
compared to human MSCs.
2. Genetic Manipulation of Mesenchymal Stem Cells
Also, a major limitation to the efficacy of mesenchymal stem cell therapy is the
poor viability of the transplanted cells. To date, the functional improvement
from mesenchymal stem cell therapy has been modest, and genetic modification
162 Chen LB, Jiang XB, Yang L. Differentiation of rat marrow mesenchymal stem cells into pancreatic islet beta-cells. World J Gastroenterol 2004; 10(20): 3016-3020. 163 Gorba T, Allsopp TE. Pharmacological potential of embryonic stem cells. Pharma Research 2003; 47(4): 269-278.
88
of stem or progenitor cells may represent an important strategic advancement in
regenerative medicine. 164
By combining gene with cell therapy, scientists may be able to enhance stem cell
function and viability. There is evidence that genetic modification can improve
survival, metabolic characteristics, contractility, proliferative capacity, or
differentiation of the stem cells. Furthermore, such cells may become a vehicle
for gene therapy whose secreted gene products can exert paracrine or endocrine
actions that may result in further therapeutic benefits.
One of the early groups to conceptualize this approach was Dr. Mangi and his
collaborators at Cleveland Clinic Foundation in Cleveland, OH. His group showed
that MSCs over-expressing the anti-apoptotic gene Akt1 (Akt-MSCs) became
more resistant to apoptosis in vitro and in vivo.165 Furthermore, when injected
into infarcted hearts, Akt-MSCs dramatically limit ventricular remodeling and
improve cardiac function.
Another group pursuing this concept is Tang et al. of the Keck Graduate Institute
of Applied Life Science in Claremont, CA. They showed that a higher number of
mesenchymal stem cells transduced with heme oxygenase (HO)-1 survive when
they are injected into infarcted hearts, leading to more efficient healing
compared with control MSCs.166
In research situations in which genetically manipulated cells must be introduced
to a host, animal models are typically preferred, because FDA approval to initiate
clinical trials in humans is usually a limiting factor. As a result, rat mesenchymal
stem cells have value as tools for testing the effects of gene therapy induced
through knock-out, knock-down, or up-regulating manipulation and performed
to improve stem cell viability or other characteristics. 164 Dzau VJ, Gnecchi M, Pachori AS. Enhancing Stem Cell Therapy Through Genetic Modification. J Am Coll Cardiol 2005; 46:1351-1353. 165 Mangi AA, Noiseux N, Kong D, et al. Mesenchymal stem cells modified with Akt prevent remodeling and restore performance of infarcted hearts. Nat Med 2003; 9: 1195-1201. 166 Tang YL, Tang Y, Zhang YC, Qian K, Shen L, Phillips MI, et al. Improved graft mesenchymal stem cell survival in ischemic heart with a hypoxia-regulated heme oxygenase-1 vector J. Am Coll Cardiol 2005; 46: 1339-1350.
89
3. Potential for Genetically-Engineered Mesenchymal Stem Cell Reporter Lines
Also, as the stem cell industry matures, companies such as Invitrogen, Inc., are
moving into the development of genetically engineered stem cell lines.
Currently, Invitrogen is developing novel engineered human embryonic stem cell
(hESC) reporter lines. The company is accomplishing this by pairing promoters
and reporter elements to create cell lineage “beacons” that can be used to
screen small molecules, potential regulatory proteins, or noncoding RNAs for
their impact on cell differentiation.167 This approach will ultimately be used to
manage cell differentiation toward a specific lineage for a therapeutic
application by expressing or silencing genes of interest.
Differentiation of ES cells can then be exploited to develop powerful screens for
identifying drugs that induce the body's own power of regeneration to repair
and activate cells that are needed to treat common disorders, such as diabetes
and cardiovascular and neurodegenerative disease.
As of now, this technique of generating stem cell reporter lines has not been
performed with mesenchymal stem cell lines, human or mouse. However, this is
an interesting application that is likely to be applied to mesenchymal stem cells
in the future, with Invitrogen already expressing interest in the possibility.
Indeed, the screening of small molecules, regulatory proteins, and noncoding
RNAs for their impact on cell differentiation acts to support the development of
drugs that can induce the body’s own power of regeneration. Because
mesenchymal stem cells are now in clinical trials for several different
applications, their potential for treating serious diseases is being recognized, and
more tools for exploring their potential are likely to be developed, including cell
reporter lines.
167 Invitrogen, Inc. Available at: www.invitrogen.com [Accessed December 1, 2010].
90
XII. SERUM-FREE SUPPLEMENTS FOR USE IN MSC RESEARCH
MSCs are currently in use for therapeutic applications, but until now, their growth in
vitro has required the use of media containing animal-derived serum. Serum-based
media includes undefined elements that can make performance vary from lot-to-lot. In
addition, the inclusion of undefined animal components can raise questions with
respect to regulatory agency approval of therapeutics based on MSCs.
The proposed advantages of using serum-free supplements for MSC research include:
Superior expansion efficiency
Better quality hMSCs
Better lot-to-lot consistency
At this time, there is market demand for serum-free supplements qualified for use with
MSCs, but relatively few commercial suppliers.
A. Invitrogen
At this time, the primary company providing a serum free supplement for MSCs is
Invitrogen. Invitrogen offers STEMPRO® MSC SFM, the first serum-free medium
(SFM) specially formulated for the growth and expansion of human mesenchymal
stem cells. It was launched on May 22, 2008.168 In a strategic decision, Invitrogen
also launched two kits designed to allow the differentiation of human MSCs to either
adipocytes or osteocytes at the same time.
STEMPRO® MSC SFM is completely defined, meaning that each element is present in
the media at a known quantity, thereby eliminating significant lot-to-lot variability.
The media maintains MSCs in an undifferentiated state for more than nine passages,
168 Invitrogen Launches the First Ever Serum Free Media for Mesenchymal Stem Cells – Invitrogen Press Releases – Business Wire. Available at: http://www.smartbrief.com/news/aaaa/industryBW-detail.jsp?id=5A915FCC-963F-4A06-BF8F-34BBBAA94E0B [Accessed March 08, 2012].
91
whereas cells in traditional media supplemented with serum start to demonstrate
diminished cellular expansion and differentiation after five passages.
Invitrogen’s STEMPRO® MSC SFM is advertised as offering:169
Superior efficiency of hMSC expansion at high cell densities — less medium,
surface area, and time
Better quality cells — primitive phenotype, retained hMSC surface marker
expression, normal gene expression profiles, and maintained CFU-F and tri-
lineage mesoderm differentiation potential beyond passage five
Batch-to-batch consistency with each lot—produced under cGMP and
qualified using a hMSC performance assay
No or little adaptation required from serum-supplemented medium
At this time, Invitrogen does not offer STEMPRO® MSC SFM designed for use with
mouse MSCs, but this may be a product that it will make available in the near future.
B. Celprogen
Interestingly, Celprogen advertises on its website that it provides Rat Mesenchymal
Bone Marrow Serum Free Media (Cat. No. M55096-23) for USD $125. However, it is
nearly impossible to discover the existence of this product using the search phrase
“serum free media,” or related queries, on any of the major search engines (Google,
Yahoo, and Ask), which dominate over 95% of all search activity.
Essentially, the only way to find this product is to visit the Celprogen website
(www.celprogen.com) and then use the intra-site search tool to search for “serum-
free media.” Also, the link on the Celprogen website that would allow a visitor to
view their Product Sheet for the Rat Mesenchymal Bone Marrow Serum Free Media
is broken. Thus, it is highly unlikely that Celprogen is generating sales from this
product, since purchasing it is a very user “unfriendly” process.
169 STEMPRO® MSC Product Page – Invitrogen, Inc. – Corporate Website. Available at: http://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/Cell-Culture/Stem-Cell-Research/Stem-Cell-Research-Misc/StemPro-MSC-SFM.html [Accessed December 1, 2010].
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Celprogen also advertises on its website that it offers Human Mesenchymal Bone
Marrow Serum Free Media (Model # M36094-21) and Human Mesenchymal Bone
Marrow Serum Free Differentiation Media (M36094-21D). While the Product Sheets
for these products are active links, it is again nearly impossible to discover the
existence of these products using the search phrase “serum free media,” or related
queries, on any of the major search engines (Google, Yahoo, and Ask).
Based on interviews with researchers actively using serum-free supplements, it is
believed that Celprogen’s product quality is unpredictable.170 Some interview
subjects were unhappy with products purchased from them in the past. While
Celprogen is an innovative company in some respects, numerous product data
sheets have been removed from their website in the past 12 months. Consequently,
uneasy Celprogen consumers have brought their business elsewhere.
C. Millipore
Millipore also offers a serum-free medium called HEScGRO, but it is optimized for
work with embryonic stem-cell lines, rather than mesenchymal stem cell lines.
Millipore’s HEScGRO medium is the first commercially available, animal component-
free (ACF) medium that has been successfully tested for human embryonic stem cell
culture. 171 Millipore’s hES cell medium is designed to support the growth and
expansion of undifferentiated human ESCs and is specially formulated to meet the
special requirements of human embryonic stem cell culture. It has been successfully
tested and proven to maintain the pluripotent nature of several hES cell lines
including MEL-1, MEL-2, and H1. This medium is defined, serum-free, and animal
component- free, and does not require additional supplementation to maintain cells
in their pluripotent state.
170 BioInformant, Investigational Research Team. Direct Communication: Telephone Interviews. March 06, 2012. 171 Millipore’s HEScGRO Medium Advances Human Embryonic Stem Cell Research – Millipore Press Release. Available at: http://www.millipore.com/pressroom/pr3/hescgro [Accessed March 09, 2012].
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While this is not a product that Trevigen will want to license from another
manufacturer for use with its Rat Mesenchymal Stem Cell products, it does show
that Millipore recognizes the importance of serum-free medium and is developing
products to meet this market need. As such, they may launch a serum-free product
for mesenchymal stem cells in the near future.
D. STEMCELL Technologies
At this time, STEMCELL Technologies does not offer serum-free medium for rat
mesenchymal stem cells (MSCs). However, the company does offer StemSpan®
Serum-Free Expansion Media (SFEM) that has the ability to maintain, expand, and
assay hematopoietic stem cells and progenitors.172 The company’s StemSpan® SFEM
and StemSpan® H3000 products are animal-free mediums containing only human-
derived or recombinant human proteins.173 The company also offers NeuroCult™
Proliferation Media, a standardized, serum-free NeuroCult™ Proliferation Media for
the expansion of neural stem and progenitor cells in neurosphere and adherent
monolayer cultures, with optimized formulations for human, mouse and rat cells
from CNS tissues.174
As a self-described “leading supplier of serum substitutes and serum-free media that
support the maintenance and expansion of hematopoietic stem cells and
progenitors,” it seems likely that STEMCELL Technologies will develop serum-free
medium for mesenchymal stem cells in the near future, if they are able to
successfully license or circumvent the patent restrictions described in the following
section.
172 STEMCELL Technologies, Corporate Website. “StemSpan® Serum-Free Expansion Media” Page: http://www.stemcell.com/en/Technical-Resources/c5f87/StemSpan-Serum-Free-Expansion-Media.aspx?id={C5F87ADE-DFF6-4E25-9D8C-9764A72483A8}. [Accessed March 10, 2012]. 173 STEMCELL Technologies, Corporate Website. “StemSpan® SFEM Product Page” Page: http://www.stemcell.com/en/Products/All-Products/StemSpan-SFEM.aspx. [Accessed March 9, 2012]. 174 STEMCELL Technologies, Corporate Website. “NeuroCult™ Proliferation Media” Page: http://www.stemcell.com/en/Technical-Resources/76a0a/NeuroCult-Proliferation-Media.aspx?id={76A0A565-7EC8-49C2-9DA7-C733A737FA78}. [Accessed March 10, 2012].
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E. United States Patent 7109032: Serum-free medium for mesenchymal stem cells
United States Patent 7109032 is entitled, “Serum-free medium for mesenchymal
stem cells.” It was published on 09/19/2006 and is held by:
Consorzio per la Gestione del Centro di Biotecnologie Avanzate (Genoa, IT),
and
Istituto Nazionale per la Ricerca Sul Cancro (Genoa, IT)
The abstract for this patent reads:
Serum-free media for growth and proliferation of chondrocytes and
mesenchymal stem cells in culture are provided. A serum-free medium for growth
of chondrocytes includes a serum-free composition comprising FGF-2, linoleic
acid, ascorbic acid, B-mercaptoethanol, transferrin and dexamethasone. Further,
the composition comprises EGF, PDGFbb, insulin and albumin. A method for
growing chondrocytes in a serum free medium comprising the composition is also
provided. Also provided for mesenchymal stem cell growth, is a serum-free
medium which includes a composition comprising FGF-2, LIF, SCF, pantotenate,
biotin and selenium.
For any research supply companies interested in producing their own serum-free
medium for use with MSCs, it will be necessary to either contact the assignees of
this patent to get express permission, alternatively, it will be necessary to creatively
circumvent the claims of this patent.
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XIII. END-USER PREFERENCES (SCIENTIST PANEL)
The approach taken to survey end-user preference with regard to MSC research product
preferences was to implement an electronically-based end-user survey. In May 2012,
the survey questions below were distributed to stem cell scientists. 147 completed
surveys and 16 partially completed surveys were received as the final pool of
respondents.
An electronically-based survey allowed for a larger geographical audience than could
have been reached using a focus group. The results were able to integrate US-based
researchers, as well as a global audience that included respondents from the Canada,
the U.K., Europe, China, Japan, and Australia.
A. Preferred Tissue Source
The percentages below represent the percent of respondents who reported use of
the following source as their “primary and preferred’ tissue source when involved
with mesenchymal stem cell research.
TISSUE SOURCE % OF RESPONDENTS
Bone Marrow 54.1
Umbilical Cord Blood 19.5
Adipose Tissue 11.8
Dental Pulp 1.9
Liver 0.3
Pancreas 0.1
All Other Sources 12.3
TOTAL = 100.0%
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0.1%0.3%
1.9%
11.8%
12.3%
19.5%
54.1%
Preferred Tissue Source
Pancreas
Liver
Dental Pulp
Adipose Tissue
Other
Umbilical Cord Blood
Bone Marrow
B. Preferred Species Source
The percentages below represent the percent of respondents who reported use of
the following MSC species as their “primary and preferred’ species source when
involved with mesenchymal stem cell research.
SPECIES SOURCE % OF RESPONDENTS
Human 43.0
Rat 30.5
Mouse 19.9
Pig 1.8
Sheep 0.4
Canine 0.2
Goat 0.1
Other 4.1
TOTAL = 100.0%
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C. MSC Product Preferences
The scientist panel was also presented with a list of mesenchymal stem cell product
categories and asked to rank them from “High Utility” (1st Rank) to “Low Utility” (10th
Rank). Across the population of respondents, the following results were received,
presented below. The percent displayed to the right of each product category
represents the percent of respondents who ranked it as the single product category
that represented the highest perceived utility to them (1st rank).
98
RANK MSC PRODUCT CATEGORY % OF RESPONDENTS
1st Primary Mesenchymal Stem Cells 29.1
2nd MSC Differentiation Kits 22.7
3rd (T) MSC-Qualified Cell Culture Products 12.2
3rd (T) MSC Antibodies 12.0
5th MSC-Specific Growth Factors, Chemokines and Cytokines 9.3
6th Differentiation-Inducing Medium and Supplements 4.3
8th (T) Extracellular Matrix Proteins 3.1
8th (T) MSC Expansion Kits 3.0
8th (T) Serum-free and Specialty Media 2.3
10th Transfection Kits 2.0
TOTAL = 100.0
29.1%
22.7%12.2%
12.0%
9.3%
4.3%3.1%
3.0%
2.3%2.0%
MSC Product Preferences
Primary MSCs
MSC Differentiation Kits
MSC-Qualified Cell Culture Products
MSC Antibodies
MSC-Specific Growth Factors, Chemokines and Cytokines
Differentiation-Inducing Medium and Supplements
Extracellular Matrix Proteins
MSC Expansion Kits
Serum-free and Specialty Media
Transfection Kits
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D. Most Commonly Reported Antibodies
Because many research supply companies are interested in scientist preferences
regarding antibodies to stem cell markers, this section sought to identify the stem
cell markers most frequency used by mesenchymal stem cell researchers. To do this,
survey respondents were asked to choose from a panel of 125 potential
mesenchymal stem cell markers to indicate which ones they use in their own
research. Survey respondents were also allowed the opportunity to “write in” the
names of other stem cell markers which were not presented within the panel of
choices. However, no “write-ins” appeared frequently enough in the pool of
responses to be highlighted here.
The percentages below represent the respondents who reported utilizing the
following MSC markers in their own research within the past six months. (Note that
because the scientists were allowed to report usage of as many MSC markers as they
used regularly, the percentages sum to greater than 100%.)
ANTIBODY PERCENT OF SURVEY RESPONDENTS
ICAM-1 38.0
Collagen 1 34.0
Thy-1 33.0
VCAM 33.0
Fibronectin 31.0
Endoglin 27.0
PECAM-1 21.0
SSEA-1 19.0
CD105 13.0
TNF-Receptor 12.0
Musulin 11.0
HCAM 9.0
Transferrin Receptor 9.0
CD44 6.0
LCA 5.0
Blast 1 4.0
Integrin B1 2.6
STRO-1 2.6
Nucleostemin 1.0
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E. Terms Used in Online Product Search
In addition, the scientist panel was asked to report search terms that they used
within the past month to perform online (Web-based) searches for mesenchymal
stem cell products. Each participant was allowed up to five responses, and no
suggestions were provided. Responses were entered into a blank text box. Overall,
more than 45 different search terms were received back from the pool of
respondents, a surprisingly large number and one that shows the diversity of
thought that can occur across a population of researchers.
However, the top ten search terms used to perform online product searches were
reported with high frequencies. These terms are listed below, in decreasing order
from most common to least common:
38.0
34.0
33.0
33.0
31.0
27.0
21.0
19.0
13.0
12.0
11.0
9.0
9.0
6.0
5.0
4.0
3.0
3.0
1.0
0.0 5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0
ICAM-1
Collagen-1
Thy-1
VCAM
Fibronectin
Endoglin
PECAM-1
SSEA-1
CD105
TNF-Receptor
Musulin
HCAM
Transferrin Receptor
CD44
LCA
Blast 1
Integrin B1
STRO-1
Nucleostemin
Preferred Antibody
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RANK SEARCH TERM
1 Mesenchymal Stem Cell
2 Mesenchymal Stem Cells
3 MSC Antibody
4 MSC Ab
6 Mesenchymal Stem Cell Kit
5 Mesenchymal Ab
8 MSC Kit
7 Mesenchymal Stem Cell Antibody
9 MSC
10 Mesenchymal Research Product
There are several interesting findings from this list, summarized below:
The only difference between the first and second most common search
terms was use of singular, versus plural, description.
Surprisingly, four mesenchymal stem cell antibody terms made this list,
suggesting frequent search for this product type.
Interestingly, no MSC-specific cell culture terms made this list. (The search
term “Mesenchymal Cell Culture” ranked 16th overall and was the first of this
type.)
Researchers use the abbreviations “MSC” and “Ab” with relatively high
frequencies in online search.
Next, a comparative analysis was done to determine whether research supply
companies investing money into online advertising using the Google Adwords
program are aligning decision-making in a fashion so as to get greatest benefit from
online product searches being performed within the research community. To
evaluate this, the search terms above were screened against Google Adwords results
(Companies and Bid Amount175) and the findings are presented in the table below.176
175 “Bid amount” is the amount of money a company is willing to pay for a specific search term. Daily Google Adwords budget and bid amount for a specific search term determine a company’s Google Adwords placement in response to user search. As such, bid
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TABLE: Top MSC Search Terms Used by Scientific Researchers and Relative Competitiveness for Google Adwords Placement
RANK SEARCH TERM
# OF RESEARCH SUPPLY
COMPANIES PAYING FOR
SEARCH TERM (GOOGLE
ADWORDS)
SPECIFIC COMPANIES PAYING FOR SEARCH TERM
(LISTED FROM HIGHEST-TO-LOWEST BID RATE)
1 Mesenchymal Stem Cell 3
Stem Cell Technologies, Trevigen, Miltenyi Biotec
2 Mesenchymal Stem Cells 7
Lonza, Stemgent, Promocell, Trevigen, ScienCell, Assay Designs, Miltenyi
Biotec
3 MSC Antibody 7
BD Biosciences, Epitomics, R&D Systems, Genscript, Abnova,
Stemgent, Miltenyi Biotec
4 MSC Ab 0 None
6 Mesenchymal Stem Cell Kit 7
Genscript, Stem Cell Technologies, Lonza, Miltenyi Biotec, R&D Systems,
Promocell, Trevigen
5
Mesenchymal Ab 8
Stem Cell Technologies, Trevigen, Novus Bio, Cell Sciences, Promocell,
Lonza, SDIX, ScienCell
8 MSC Kit 0 None
7 Mesenchymal Stem Cell Antibody 7
Genscript, eBioscience, BD Biosciences, R&D Systems, Epitomics,
Miltenyi Biotec, Stem Cell Technologies
9 MSC 0 None
10 Mesenchymal Research Product 8
Open Bio, Stem Cell Technologies, Genscript, Promocell, Prosci Inc,
Paragon Bioservices, Trevigen, SDIX
amount reflects the degree to which a company prioritizes a specific search term, as well as the overall size of that company’s Google Adwords budget. 176 Google Adwords, “Google Adwords Tool” for Keyword Analysis: https://adwords.google.com/o/Targeting/Explorer?__c=1000000000&__u=1000000000&ideaRequestType=KEYWORD_IDEAS. Ten (10) Search Term Analysis. Executed March 15, 2012.
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There are several interesting findings from this comparison, summarized below:
First, a number of the top search terms do not have any research product
companies bidding for them. These search terms are highlighted in yellow in
the table above. They represent excellent opportunities for research product
companies to capture search traffic.
Several of the companies advertising for the top search terms link to general
stem cell research product pages and do not offer truly MSC-specific
products.
The top search term “Mesenchymal Stem Cell” does not have nearly as many
Adword competitors (three) as several of the lower ranking search terms (for
which there are seven to eight competitors, in some cases).
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XIV. RESEARCH PRODUCT OPPORTUNITIES & SUGGESTIONS
The following are novel research product opportunities and suggestions for companies
that wish to offer competitive stem cells products to the rapidly growing, well-funded
MSC research community. The suggestions here are intended to represent unique
market opportunities and unfulfilled niches, rather than be a comprehensive listing of
potential product offerings.
A. Rat MSCs Derived from Synovium
Several companies already offer primary mesenchymal stem cells (usually
derived from bone marrow or adipogenic tissue), as well as kits that include
ready-to-use mesenchymal stem cells as a component. However, there is an
interesting market opportunity to offer variations of these products.
Specifically, there are known benefits associated with MSCs derived from
synovium, but to date, there is only a single commercial provider of synovium-
derived mesenchymal stem cells, Thermo Scientific, which offers “Hyclone CET
Human Amniotic Mesenchymal Stem Cells.”
Interesting research was performed by Yoshiumra, et al., published in Cell
Tissue Research in March 2007.177 Yoshiumra’s research, titled “Comparison of
rat mesenchymal stem cells derived from bone marrow, synovium, periosteum,
adipose tissue, and muscle,” compared properties for yield, expansion, and
multipotency for rat MSCs isolated from bone marrow, synovium, periosteum,
adipose, and muscle.
As a result of this research, Yoshiumra and colleagues determined that the
colony number per nucleated cells derived from synovium was 100-fold higher
than that for cells derived from bone marrow. With regard to expansion
177 Yoshimura, et al. Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle. Cell Tissue Research, 2007.
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potential, synovium-derived cells were the highest in colony-forming efficiency,
fold increase, and growth kinetics.
The yield and proliferation potential of rat MSCs from solid tissues was much
better than those from bone marrow. In particular, synovium-derived cells had
the greatest potential for both proliferation and chondrogenesis, indicating
their usefulness for cartilage study in a rat model. An in vitro chondrogenesis
assay also demonstrated that the pellets derived from synovium were heavier,
due to their greater production of cartilage matrix, than those from other
tissues, confirming their advantage in chondrogenesis.
The Oil Red O positive colony-rate assay demonstrated higher adipogenic
potential in synovium- and adipose-derived cells.
As mentioned, there is only one commercial provider of synovium-derived
mesenchymal stem cells, Thermo Scientific, and this company offers human
synovium-derived mesenchymal stem cells. To date, no commercial provider
exists for synovium-derived rat mesenchymal stem cells, despite demonstrated
advantages of MSCs derived from this source. As such, this may represent a
unique product opportunity for research products companies to consider.
B. Novel MSC Differentiation Kits
Differentiation assays can be used to demonstrate multi-potentiality of a
starting cell population of mesenchymal stem cells.
Differentiation assays that are already commercially available for use with rat
mesenchymal stem cells include:
Millipore’s Mesenchymal Stem Cell Adipogenesis Kit (Cat. No. SCR020):
Millipore's Mesenchymal Stem Cell Adipogenesis Kit contains reagents
that readily differentiate rat bone marrow-derived mesenchymal stem
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cells to an adipogenic lineage, as assessed with Oil Red O staining of lipid
vacuoles in mature adipocytes.
Millipore’s Mesenchymal Stem Cell Osteogenesis Kit (Cat. No. SCR028):
Millipore's Mesenchymal Stem Cell Osteogenic Kit contains reagents that
readily differentiate rat bone marrow-derived mesenchymal stem cells
to an osteogenic lineage, as assessed by Alizarin Red staining.
AND
Trevigen’s Mesenchymal Stem Cell Adipogenic Differentiation Kit (Cat. No.
5010-024-K): A differentiation kit that follows traditional methods of
adipogenic differentiation, by growing MSC in medium supplemented with
insulin, isobutyl methyl xanthine (IBMX), indomethacin, and dexamethasone.
Trevigen’s Mesenchymal Stem Cell Osteogenic Differentiation Kit (Cat. No.
5011-024-K): A differentiation kit that contains reagents optimized to direct
mesenchymal stem cells grown on Cultrex® Rat Collagen I to undergo
osteogenic differentiation in a defined growth medium supplemented with
ascorbic acid, ß-glycerol phosphate, dexamethasone, and qualified fetal
bovine serum.
At this time, Millipore and Trevigen are the only companies that offer
Adipogenic and Osteogenic Kits for use with rat mesenchymal stem cells within
the US. Millipore launched its rat MSC differentiation kits in 2007, and Trevigen
launched its in 2009. However, several other companies, such as Thermo
Scientific and Invitrogen, offer Adipogenic and Osteogenic Kits for use with
human mesenchymal stem cells. Cyagen offers Adipogenesis and Osteogenesis
Kits for use with rat mesenchymal stem cells, but only distributes its products
within China.
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Note that Chondrogenic and Neural Differentiation Kits designed to support
differentiation of rat mesenchymal stem cells into these lineages are not yet
commercially available. As such, research product companies developing MSC
products could consider providing these kits.
Differentiation assays that are commercially available, but designed for use
with human MSCs include:
Thermo Scientific’s HyClone AdvanceSTEM™ Osteogenic Differentiation
Kit: This kit was developed to support the differentiation of human
mesenchymal stem cells into an osteogenic lineage.
Thermo Scientific’s HyClone AdvanceSTEM™ Adipogenic Differentiation
Kit: This kit was developed to support the differentiation of human
mesenchymal stem cells into an adipogenic lineage.
Thermo Scientific’s HyClone™ AdvanceSTEM™ Neural Differentiation Kit:
This kit was developed to support the differentiation of human
mesenchymal stem cells into a neural lineage.
Note that there is not yet a commercially available Chondrogenic
Differentiation Kit designed to support differentiation of human
mesenchymal stem cells into this lineage.
Also, the following are differentiation kits that are not yet commercially
available to support the differentiation of either human or rat-derived MSCs:
Myocardial Differentiation Kit: Mesenchymal stem cells have
demonstrated the potential to replace infarct scar in rat myocardial
infarction models. Specifically, allogeneic MSCs injected into rat
infarcted myocardium survive as long as six months and express markers
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suggesting cardiac muscle and endothelium phenotypes.178 A kit
containing reagents to readily differentiate rat bone marrow-derived
MSCs toward a myocardial lineage would be an interesting and unique
product for research product companies to consider developing.
Similarly, a Myocardial Differentiation Kit to support differentiation of
human MSCs into a myocardial lineage could be created.
Hepatocytic Differentiation Kit: Rat mesenchymal stem cells from bone
marrow have demonstrated the potential to differentiate towards
hepatocytic lineage, with the microenvironment playing a decisive role
in the induction of hepatic differentiation of rMSC.179 A kit containing
reagents to readily differentiate rat bone marrow-derived MSCs toward
a hepatocytic lineage would be another novel differentiation kit for
research product companies to consider for development.
Similarly, a Hepatocytic Differentiation Kit to support the differentiation
of human MSCs into a hepatocytic lineage could be created.
C. Mesenchymal Stem Cell Expansion Media
A less exciting but practical opportunity would be to offer mesenchymal stem
cell expansion media, as there are currently relatively few commercial
providers for this type of expansion medium compared to other cell types.
Possibilities in this area include:
DMEM-low glucose, without glutamine
10% heat-inactivated fetal bovine serum
2 mm L-Glutamine
1X solution of Penicillin and Streptomycin
178 Dai, et al. Allogeneic Mesenchymal Stem Cell Transplantation in Postinfarcted Rat Myocardium. The Heart Institute, Good Samaritan Hospital 2005. 179 Lange, et al. Liver-specific gene expression in mesenchymal stem cells is induced by liver cells. World Jour of Gastroenterol 2005. Available at: http://www.wjgnet.com/1007-9327/11/4497.pdf [Accessed March 14, 2012].
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D. Antibodies to Mesenchymal Stem Cell Markers
Research product companies can also consider offering antibodies to positive
and negative cell markers of MSCs, as there are currently few commercial
providers of MSC antibodies relative to other cell types, and isolation of
mesenchymal stem cells from primary tissue remains restricted by the limited
selectivity of available markers.180
To date, CD271 is one of the most specific markers for bone marrow-derived
MSC. Additionally, CD105 (SH2), CD73 (SH3/4), and STRO-1 represent positive
MSC markers leading in popularity and usage, while CD34, CD45, and CD14
represent negative MSC markers leading in popularity and usage.181
Also, it has been recently proposed that each of the following markers are also
suitable for the isolation of highly purified MSCs: platelet-derived growth factor
receptor-β (CD140b), HER-2/erbB2 (CD340), frizzled-9 (CD349), W8B2 antigen,
W1C3, W3D5, W4A5, W5C4, W5C5, W7C6, 9A3, 58B1, F9-3C2F1, and HEK-3D6.61
Based on findings by Buhring et al., these MSCs may have the potential to be
used as an improved starting population for transplantation in cartilage repair,
myocardial infarction, and diseases like osteogenesis imperfecta,.
As MSC research progresses, there will continue to be newly identified positive
and negative markers for MSCs. Competitive research supply companies should
considering offering antibodies that include: 1) highly specific markers
(example: CD271); 2) frequently used markers; and 3) novel, recently identified
markers. Supplying antibodies for markers within each of the above categories
will result in fulfilling a cross-section of market needs.
180 Buhring HJ, et al. Novel Markers for the Prospective Isolation of Human MSC. NY Acad of Science 2007; 1106: 262-271. 181 BioInformant Worldwide, LLC, Investigational Research Team. Data Analysis Division. January 2012. Sources utilized include PubMed and Highwire Press Data, CRISP Grant Database, and Web Queries for Mesenchymal Stem Cell (MSC) research products within the trailing 12 months.
110
Additionally, the following are possible antibodies to offer for the purposes of
identifying positive and negative cell markers for Rat MSCs:
Positive Cell Markers for Rat MSCs:
Antibody to Beta-1
Antibody to CD54
Negative Cell Markers for Rat MSCs:
Antibody to CD14
Antibody to CD45
E. MSC Growth Factors
To optimally expand MSCs in vitro, a combination of mitogenic factors are required,
including:182
Platelet Derived Growth Factor [PDGF]
Epidermal Growth Factor [EGF]
basic Fibroblast Growth Factor [bFGF]
Transforming Growth Factor-ß [TGFß]
Insulin Like Growth Factor [IGF]
Also, an MSC culture system that incorporated Platelet Derived Growth Factor-BB
[PDGF-BB] in addition to EGF was shown to result in a cell doubling time of 48 to 72
hours and an ability to support purified MSCs for more than 60 population
doublings.183
Thus, research supply companies could also consider offering recombinant proteins
suitable for culture of MSCs.
182 Kuznetsov SA, Friedenstein AJ, Robey PG. Factors required for bone marrow stromal fibroblast colony formation in vitro. Br J Haematol 1997; 97(3): 561-70. 183 Reyes. Purification and ex vivo expansion of postnatal marrow mesodermal progenitor cells. Blood 2001; 98(9): 2615-2625.
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F. Monoclonal Antibodies and Kits for Phenotyping MSCs
As mentioned previously, MSC populations are typically heterogeneous in nature
and present with variable expression of cell surface markers. There is no single test
that can be performed on a cell to determine whether that cell is an MSC. There are
surface antigens that can be used to isolate a population of cells with similar self-
renewal and differentiation capacities, but MSCs, as a population, generally do not
all express the proposed markers, and it is not certain which ones must be expressed
in order for a cell to be definitively identified as an MSC.
Regardless, the ability to correctly identify and sub-stratify mesenchymal stem cell
phenotypes will be crucial for their use in regenerative therapies. For this reason,
there is growing market demand for approaches to accurately phenotype MSCs
based on either single or collections of cell surface molecules.
Antibodies recognizing certain cell surface molecules are probably the best reagents
to characterize and isolate MSCs of various phenotypes. To date, a number of MSC
phenotypes have been reported, including but not limited to: CD271+, Sca-1+,
CD29+, CD44+, c-Kit+, CD105+, CD45–, CD73+, STRO-1+, CD31+184 and Sca-1+,
CD29+, CD44+, CD81+, CD106+, Nucleostemin+ and CD116–, CD34–, CD45–, CD48–,
CD117–, CD14- and CD135–.185
While most of these antibodies are commercially available as independent products,
few companies are currently offering sets of these markers in panel or kit formats
for detailed phenotyping of MSC populations. As MSC therapy research progresses
and more treatments enter clinical testing stages, detailed phenotyping will increase
in both frequency and popularity.
184 Sun, et al. Isolation of mouse marrow mesenchymal progenitors by a novel and reliable method. Stem Cells 2003; 21(5): 527-535. 185 Baddoo, et al. Characterization of mesenchymal stem cells isolated from mouse bone marrow by negative selection. Jour of Cell Biochem 2003; 89(6): 1235-1249.
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G. Chips for Microarray Analysis of Gene Expression in MSCs
Finally, research product companies can also consider the potential for offering
chips for microarray analysis of gene expression in mesenchymal stem cells. These
chips could be general MSC expression chips or chips designed for analysis of MSCs
that have differentiated into specific mesenchymal lineages, such as adipogenic
cells, osteogenic cells, or chondrogenic cells.
As mentioned earlier, PubMed is a service of the US National Library of Medicine
that is a meta-database of scientific publications. The charts below analyze PubMed
publication rates for the terms: 1) “Mesenchymal” and “Microarray,” and 2)
“Mesenchymal” and “Gene Expression.”
TABLE: PubMed Analysis of Publication Rates by Year (Search Term "Mesenchymal" and "Microarray")
YEAR # OF PUBLICATIONS
2004 30
2005 42
2006 64
2007 76
2008 97
2009 131
2010 142
2011 151
TABLE: PubMed Analysis of Publication Rates by Year (Search Term "Mesenchymal" and "Gene Expression")
YEAR # OF PUBLICATIONS
2004 333
2005 430
2006 512
2007 613
2008 613
2009 667
2010 713
2011 778
113
Impressively, the data set for the search terms “Mesenchymal” and “Microarray”
shows an approximately five-fold increase in microarray analysis of mesenchymal
cells over the past five years. The data set for the search terms “Mesenchymal” and
“Gene Expression” shows a doubling over the past five years. These data sets were
generated in order to show that there has been a trend toward increased interest in
profiling the gene expression of mesenchymal stem cells.
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XV. POTENTIAL CUSTOMERS: INDIVIDUAL LABS AND END-USERS OF MSC RESEARCH
PRODUCTS
Another topic of interest to research supply companies developing stem cell product
lines is how to design effective communication strategies for accessing the marketplace.
For vendors of research supplies to generate substantial revenue from sale of MSC
research products, they must be able to effectively communicate with scientists
involved in the study of stem cells. To that end, the table and graph below detail the
number of researchers in BioInformant’s database who are known to be currently
conducting mesenchymal stem cell research.
Country
# of Researchers In Our Database Working With Mesenchymal Stem Cells
% of World's Total MSC Research
USA 36,921 21.1%
India 21,893 12.5%
Japan 19,058 10.9%
Australia 17,058 9.7%
Germany 11,960 6.8%
China 8,894 5.1%
United Kingdom 8,820 5.0%
Italy 6,929 4.0%
Canada 6,538 3.7%
South Korea 3,014 1.7%
France 2,527 1.4%
Brazil 2,482 1.4%
The Netherlands 2,189 1.2%
Russia 1,196 0.7%
Sweden 1,129 0.6%
Other 24,632 14.1%
TOTAL 175,240 100.0%
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Additionally, highlighted below are individual labs and end-users of mesenchymal
research products that have exhibited unusually prolific amounts of mesenchymal stem
cell research, over a trailing three-year interval.
A. General MSC Research Labs
Due to the vast size of the mesenchymal stem cell research community, it is not
possible to present all labs currently conducting MSC research. Therefore, this section
highlights select labs and research institutions that are conducting prolific
mesenchymal stem cell research, as determined by either an: 1) Increasing rate in
MSC publications of 25% of more per year over the trailing 2-year interval (January 1,
2010 through December 31, 2011);186 or 2) Twenty or more active MSC researchers
186 Calculated using screen of Pubmed Database (http://www.ncbi.nlm.nih.gov/pubmed) for the search terms: ["Mesenchymal Stem Cell" OR "Mesenchymal Stem Cells"] OR ["Marrow Stromal Cell" OR "Marrow Stromal Cells"] OR ["Multi-potent Stromal Cell" OR "Multi-potent Stromal Cells"] OR ["Colony-Forming Unit-fibroblast" OR "Colony-Forming Unit-fibroblasts"]. Screen encompassed four technical terms for the cell type, as well as variations in singular versus plural usage of terminology.
21.1%
12.5%
10.9%
9.7%6.8%
5.1%
5.0%
4.0%
3.7%1.7%
1.4%
1.4%
1.2%
0.7% 0.6%
14.1%
MSC Researchers USA
India
Japan
Australia
Germany
China
United Kingdom
Italy
Canada
South Korea
France
Brazil
The Netherlands
Russia
Sweden
Other
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involved within a single organization.187 Contact information for these prolific MSC
research centers, including name, location, phone, and email, is provided, as available.
The California Institute for Regenerative Medicine (CIRM): CIRM is supported by funding that is intended to advance stem cell research in California, including US $822M to develop and enhance fundamental knowledge of stem cell biology, US $899M for pre-clinical research and development, and US $657M for clinical trials. Contact Information: Phone: (415) 396-9100. Email: [email protected].
Institute for Stem Cell and Regenerative Medicine, University of Southern California in Los Angeles. Contact Information: Phone: (310) 825-4958. Email: [email protected].
Tulane Center for Gene Therapy, Tulane University Health Sciences Center, 1430 Tulane Avenue, SL-99, New Orleans, Louisiana, 70112: The Tulane Center for Gene Therapy received a grant funded by the NIH to provide well-characterized human adult stem cells, rat stem cells, and mouse stem cells to academic researchers worldwide upon request.
Contact Information: Phone: (504) 988-7711. Fax: (504) 988-7710. Email: [email protected].
Whitehead Institute for Biomedical Research and M.I.T, Cambridge, Massachusetts, 02142.
Contact Information: Robert A. Weinberg. Email: [email protected].
Brigham and Women's Hospital, Department of Pathology, Boston, Massachusetts, 02115. Contact Information: Janina A. Longtine, MD. Phone: (617) 732-7444. Email: [email protected].
Dana-Farber Cancer Institute, Department of Medical Oncology, Harvard Medical School, Boston, Massachusetts, 02115. Contact Information: Elisabetta Mueller, Thomas Benjamin, and others. Phone: (617) 432-1960. Email: [email protected]
187 “Active MSC researcher” is defined as a researcher who has published one or more articles/papers on subject(s) pertaining to mesenchymal stem cells.
117
Center for Cancer Research, Department of Biology, M.I.T., 77 Massachusetts Avenue, E18-580, Cambridge, Massachusetts.
Contact Information: Jeong-Ho Hong, Michael T. McManus, Adam Amsterdam, Ralitsa Kalmukova, Phillip A. Sharp, Nancy Hopkins. Lab Phone Number: (617) 253-0609. Email (Nancy Hopkins’ Research Assistant): [email protected]
Division of Biological Engineering, M.I.T., 77 Massachusetts Avenue, E18-580, Cambridge, Massachusetts.
Contact Information: Michael B. Yaffe. E-mail: [email protected]
Department of Immunology and Infectious Diseases, Harvard School of Public Health, Harvard Medical School, Boston, Massachusetts. Contact Information: Michael B. Yaffe. E-mail: [email protected]
Department of Pathology, Harvard Medical School, 77 Louis Pasteur Avenue, Boston, Massachusetts. Contact Information: Reza Abdi, Paolo Fiorina, Chaker N. Adra, Mark Atkinson, and Mohamed Sayegh. Email (Reza Abdi): [email protected]
WiCell Research Institute in Madison, Wisconsin
Contact Information: Phone: 1-888- 204-1782. Email: [email protected]
Regenerative Medicine Institute, National Centre for Biomedical Engineering Science & Department of Medicine, National University of Ireland, Galway, Republic of Ireland.
Contact Information: J.M. McMahon, S. Conroy, M. Lyons, and others.
Institute of Clinical and Experimental Transfusion Medicine, University Medical Center Tübingen, Hoppe-Seyler-Str 3, D-72076 Tübingen, Germany.
Contact Information: Richard Schäfer, Rainer Kehlbach, Jakub Wiskirchen. Phone (Richard Schafer): +49 7071 2981665. E-mail: [email protected]
Tissue Engineering Research Group, Research Institute for Cell Engineering (RICE), National Institute of Advanced Industrial Science and Technology (AIST), 3-11-46 Nakoji, Amagasaki, Hyogo 661-0974, Japan.
Contact Information: Harumi Kagiwada, Mika Tadokoro, Noritoshi Nagaya, Hajime Ohgushi. Email: [email protected]
118
Department of Regenerative Medicine and Tissue Engineering, National Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka 565-8565, Japan. Contact Information: Akira Ohshima, Noritoshi Nagaya. Email: [email protected]
The Extracellular Matrix Research Group, Institute for Biomedical Aging Research, Austrian Academy of Sciences, Rennweg 10 A-6020 Innsbruck, Austria. Contact Information: Christine Fehrer, Gerhard Laschober, Gunter Lepperdinger. Phone: 0043 512 5839 1940. Fax: 0043 512 5839 198. E-mail: [email protected]
Laboratory of Stem Cell Research, Taiwan. Contact Information: Principal Investigator is Oscar Kuang-Sheng Lee, with assisting researchers including Szu-Ching Yvette Fang, Shu-Wen Kuo, Hui-Wen Yang, Chai-Yi Shi, and Wei-Hsien Ma. Email: [email protected]
Myeloma and Mesenchymal Research Group, at the Hanson Institute in Adelaide, South Australia. Contact Information: Andrew Zannettino and Others. Phone: +61 8 8222 3033. Email: [email protected]
Fetal Development, Stem Cells and Differentiation Group, Monash University in Australia. Contact Information: Email: [email protected]
Cell Therapy Research Group, at the University of Oslo in Norway. Contact Information: Email: [email protected]
Jane and Jerry Weintraub Center for Reconstructive Biotechnology, Jonsson Comprehensive Cancer Center, Dental Research Institute and the Division of Oral Biology and Oral Medicine. UCLA, Los Angeles, CA. Contact Information: Wendy Yang. Email: [email protected]
119
B. Osteogenic-Focused Research Labs
Skeletal Research Center, Case Western Reserve University, 2080 Adelbert Road, Cleveland, Ohio 44106.
Contact Information: Donald P. Lennon, John M. Edmison, Arnold I. Caplan, Fumiaki Sugiura, Hiroshi Kitoh and Naoki Ishiguro. Email: [email protected]
Skeletal Biology Research Center, Department of Oral and Maxillofacial Surgery, Massachusetts General Hospital and Harvard School of Dental Medicine, Boston, Massachusetts.
Contact Information: Haru Abukawa, Maria Troulis.
Trinity Centre for Bioengineering, Trinity College, Dublin, Ireland.
Contact Information: Elaine Byrne, Louise McMahon, Eric Farrell. Email: [email protected]
Science and Biotechnology Institute, Tissue Engineering and Biomaterials Laboratory, Ankara University, Ankara 06100, Turkey.
Contact Information: Aysel Koç, Nuray Emin, Eser Elçin, Y. Murat Elçin and others. Email: [email protected]
Department of Oral and Maxillofacial Surgery, Massachusetts General Hospital and Harvard School of Dental Medicine, Boston, Massachusetts.
Contact Information: Leonard Kaban.
Department of Oral and Maxillofacial Surgery, Massachusetts General Hospital, Boston, Massachusetts.
Contact Information: Roy Chuck, Williams Bradford, and others. Phone: (410) 502-1923. Fax: (443) 287-1514. Email: [email protected]
Department of Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts.
Contact Information: Shinichi Terada, Dario O. Fauza. Phone: (617) 919-2966. Fax: (617) 730-0910. Email: [email protected]
Department of Surgery, Tissue Engineering and Organ Fabrication Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts.
Contact Information: Joseph Vacanti, John E. Mayer. Email: [email protected]
120
C. Myogenic and Myocardial-Focused Research Labs
Department of Cardiothoracic Surgery and Anesthesiology, Huddinge University Hospital, Karolinska Institutet, Stockholm, Sweden.
Contact Information: KH Grinnemo , A Månsson A, and G Dellgren. Email: [email protected]
Research Centre of Heart, Brain, Hormone and Health Aging, Department of Physiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
Contact Information: Gui-Rong Li, Xiu-Ling Deng, Haiying Sun, and others. Email: [email protected]
Victor Chang Cardiac Research Institute, Sydney. ASCC Project: Cardiac Adult Stem Cells.
Contact Information: Kerry Atkinson, Dr. Gary Brooke, Dr. Tony Rossetti, Dr. Rebecca Pelekanos, and others. Email: [email protected]
D. Adipogenic-Focused Research Labs
Vascular Biology Program, Children’s Hospital Boston, Harvard Medical School, Boston, Massachusetts.
Contact Information: Ying Yu, Jasmin Fuhr, Eileen Boy, Steve Gyorffy, Shay Soker, Anthony Atala, John B. Mulliken, Joyce Bischoff. Email: [email protected]
E. Neuronal-Focused Research Labs
Department of Integrative Medical Biology, Section for Anatomy, Umeå University, Umea, Sweden.
Contact Information: Giorgio Terenghi. Phone: +44 (0)161 275 1594. Email: [email protected]
F. Immunology-Focused Research Labs
Transplantation Research Center, Brigham & Women's Hospital & Children's Hospital Boston, Harvard Medical School, Boston, Massachusetts.
Contact Information: Reza Abdi. Email: [email protected]
121
Department of Medicine, San Raffaele Scientific Institute, Milan, Italy.
Contact Information: Luigi Naldini. Email: [email protected]
Stem Cell Therapy Program, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia.
Contact Information: Abdelilah Aboussekhra, Nahed M. Hawsawi, and others. Phone: 966-1-464-7272. Fax: 966-1-442-7858. E-mail: [email protected]
Department of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, Florida.
Contact Information: Jie Deng, Bryon E. Petersen, Dennis A. Steindler, and others. Email: [email protected]
G. National Institutes of Health (NIH) Labs
National Institute of Arthritis, and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, Maryland.
Contact Information: R.S. Tuan, G. Boland, and others. Email: [email protected]
NIH/NHLBI/Division of Blood Diseases & Resources, National Institutes of Health, Bethesda, Maryland.
Contact Information: Thomas, Kelley, Fakunding, Berberich, Hunt, and others. Phone: (301) 435-0222. Email: [email protected]
Biology of Aging Program, National Institute of Aging, National Institutes of Health, Bethesda, Maryland.
Contact Information: Dr. Jill Carrington. Phone: (301) 496-6402. Email: [email protected]
Division of Blood Diseases and Resources, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland.
Contact Information: Dr. John Thomas. Phone: (301) 435-0050. Email: [email protected]
Laboratory of Stem Cell Biology, National Institutes of Health, Bethesda, Maryland.
Contact Information: Mahendra Rao, M.D., Ph.D. Phone: (301) 594-6667. Email: [email protected]
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H. Commercial Labs
Additionally, the following companies are funding internal research programs
involving the study of MSCs, for the purposes of developing therapeutic applications
or producing MSC products:
COMMERCIAL MESENCHYMAL STEM CELL LABS, 2012
United States
Name City, State
BD Biosciences San Jose, California
Cell Applications San Diego, California
Celprogen San Pedro, California
Cyagen Biosciences Sunnyvale, California
Genlantis San Diego, California
Geron Menlo Park, California
Life Technologies Carlsbad, California
Novocell San Diego, California
SA Biosciences Valencia, California
ScienCell Research Laboratories Carlsbad, California
Hemogenix Colorado Springs, Colorado
Osirius Therapeutics Columbia, Maryland
Trevigen Gaithersburg, Maryland
Abcam Cambridge, Massachusetts
Advanced Cell Technology Worcester, Massachusetts
Arteriocyte Hopkinton, Massachusetts
Genzyme Corporation Framingham, Massachusetts
Millipore Billerica, Massachusetts
Thermo Fisher Scientific Waltham, Massachusetts
R&D Systems Minneapolis, Minnesota
Lonza Allendale, New Jersey
International
Name City, Country
Cellartis Gothenburg, Sweden
CellGenix Freiburg, Germany
Miltenyi Biotec Cologne, Germany
PromoCell Heidelberg, Germany
CellMade Chappes, France
Cyagen Biosciences Guangzhou, China
Sino Biologicals Beijing, China
ES Cell International (ESI) Singapore, Singapore
LGC Standards Bangalore, India
Stempeutics Bangalore and Manipal, India
PAA Laboratories GmbH Pasching, Austria
Stem Cell Therapeutics Calgary, Alberta, Canada
Stem Cell Technologies Vancouver, British Columbia, Canada
Stempeutics Kuala Lumpur, Malaysia
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XVI. CONCLUSIONS
Literature and data concerning the biology and differentiation potential of
mesenchymal stem cells (MSCs) has become huge in less than 10 years, now totaling
nearly 13,000 publications, although some data does present as contradictory,
suggesting a heterogenic nature for MSC populations depending on source tissue,
species, and donor characteristics. MSCs appear to be an exceptionally promising tool
for cell therapy because of their unusual characteristics, which partially mimic those of
embryonic stem cells, but have advantages in terms of availability, expandability,
transplantability, and ethical implications.
Interest in therapeutic applications of human MSCs arises from their diverse ability to
differentiate into a range of cell types, as well as from their ability to migrate to sites of
tissue injury/inflammation or tumor growth. These localization properties present a
promising strategy for targeted introduction of therapeutic agents through MSC gene
therapy. In addition, MSCs possess strong immunosuppressive properties that medical
researchers are exploiting for both autologous as well as heterologous therapies.
Characterized by these properties, it not surprising that trend analysis reveals rapid
increases in research activity involving mesenchymal stem cells. Grant funding for MSC
related research has also been increasing over a trailing five-year period, although not
as rapidly. Currently, the most common types of MSC patents are those pertaining to
cell culture, cellular therapy, and differentiation, respectively. Also, the areas of muscle
repair, cartilage repair, and wound healing represent the three most common types of
MSC research, by clinical application.
Together, these indicators and more suggest that the market for mesenchymal stem cell
products will become increasingly competitive, as new participants recognize the
opportunities for growth within this market. To best understand market dynamics, this
global strategic report presents current market conditions, including scientific
publication rate data, grant rate data, patent data, and more. Other critical factors are
also explored to provide a comprehensive understanding of current industry conditions
and to identify trends within the market that could predict near-term changes.
124
With scientific publication rate increases of 112% year-over-year from 2009 to 2010,
and 116% from 2010 to 2011, mesenchymal stem cells represent a fast growing area of
stem cell research.
About BioInformant: BioInformant Worldwide, LLC, is your global leader in stem cell
industry data. As a specialty research company, we use
technology to track and identify profitable opportunities within
the stem cell industry and provide this data to companies
pursuing aggressive growth.
We are the only market intelligence company that has
specifically served the stem cell sector since it emerged, and
our singular focus allows our team to produce data that enables
you to better understand your markets, competitors, and
customers.
BioInformant has been cited by prominent news outlets that
include Nature Biotechnology, the Wall Street Journal, CBS
News, Yahoo Finance, Medical Ethics, MarketWatch, the Center
for BioNetworking, and more.
In addition, our management team comes from a BioInformatics
background – the science of collecting and analyzing complex
genetic codes – and we are trained in applying these advanced
data analysis techniques to the field of market research.
Serving Fortune 500 companies that include Pfizer, Goldman
Sachs, Beckton Dickinson, and many more, BioInformant enjoys
the status of a premium market research services provider in
the industry.
For more about our company and clients, please visit
www.BioInformant.com.
BioInformant Worldwide, LLC
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