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Manifestation of Novel Social Challenges of the European Unionin the Teaching Material ofMedical Biotechnology Master’s Programmesat the University of Pécs and at the University of DebrecenIdentification number: TÁMOP-4.1.2-08/1/A-2009-0011
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STEM CELLS AND TRANSDIFFERENTIATIONINTRODUCTION, BASIC CONCEPTS
Dr. Péter Balogh and Dr. Péter EngelmannTransdifferentiation and regenerative medicine – Lecture 1
Manifestation of Novel Social Challenges of the European Unionin the Teaching Material ofMedical Biotechnology Master’s Programmesat the University of Pécs and at the University of DebrecenIdentification number: TÁMOP-4.1.2-08/1/A-2009-0011
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Definitions
Stem cells: undifferentiated/resting cells that can divide and differentiate into mature cells of all three germ layersPluripotency: the capacity to produce several types with diverse biological characteristicsSelf-renewal: the process in which stem cells preserve their pluripotencyCommitment: the capacity of cells to restrict their differentiation spectrum/direction Differentiation: the gradual acquisition of cellular traits associated with specialization
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Stem cell research – a brief history• Early 1900’s: all blood cells come from the same
immature cells• 1950-70’s: Extensive studies on
teratocarcinomas/teratomas in mice and humans• 1963: Till and McCulloch: quantitative analysis of
hemopoietic stem cell frequency (CFU-S) • 1980-90’s: production of Tg mice from inner cell
mass of blastocysts• 1998: isolation of human stem cells from embryos:
beginning of political-ethical debate on the moral status of human embryos/stem cell research
• 2006: iPS technology begins (Yamanaka)
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TÁMOP-4.1.2-08/1/A-2009-0011Abnormal multilineage differentiation: teratoma/teratocarcinoma• Germ cell origin – testis or ovary• Mixed cell composition – malignant cells with
differentiated component (cartilage, epithelium)
• Potential use in human stem cell research in vitro
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Issues of plasticity
Plasticity: existence of parallel differentiation programs• Directionality: reversible/irreversible
(switchable), i.e. inhibition of Pax5 suspends B-cell identity; neuronal → hemopoietic transdifferentiation/switch
• Homeostasis: measured degree of commitment along various directions, responsive to external effects (altered blood cell production in infections).
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Concept of regenerative medicine
Disease model
Skin fibroblastsDiseased cell type
Pluripotent cells
PatientCell therapy
Skin punch biopsy
In vitro differentation Nuclear reprogramming
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Aims and concepts behind cellular reprogramming• Personalized cellular replacement therapy• Absent/diseased cells (genetic, degenerative,
traumatic etc. causes) and tissues corrected in a controlled cellular differentiation way
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Obstacles of cellular reprogrammingStem cell-related:• rare cell type• difficult isolation procedures• uncertain differentiation capacities
(lineage/normal/malignant)Recipient-related:• effect of previous medical treatment• problems of tissue delivery and stem cell
positioning• immunological responsiveness against the
donor cells/molecules
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Evolution of tissue regeneration• Uniform composition of early multicellular organisms
– all cells can individually regenerate• Later the ability to proliferate became restricted to a
subpopulation of the cells• Ancestral somatic stem cells (site-bound)• Migratory stem cells
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Evolution of tissue regeneration
Ancestral somatic stem cells
Somatic stem cells(migrating inside the body)
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Types of stem cells• As defined by the developmental origin:
– embryonic (neonatal)– postnatal, adult
• As defined by their physiological turnover/differentiation kinetics: – continuous, migratory (hemopoietic stem cells)– slow, sessile (liver, muscle etc)
• As defined by their differentiation spectrum:– totipotent– pluripotent– oligopotent
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Methods for reprogrammingAdvantagesDisadvantages
• Technically straightforward
• Fusion is inefficient
• Reprogrammed cells are tetraploid
Cell fusion
FibroblastES cell Tetraploid ES cell
• Indistinguishable from embryo-derived ES cells
• Technically challenging
• Sources for oocytes or zygotes
Nuclear transfer Enucleation
FibroblastOocyte or zygote ntES cell
• Technically straightforward
• Autologous to fibroblast donor
• Uses oncogenic retroviruses and transgenes
Direct reprogramming
Fibroblast iPS cell
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• Nuclear transfer: introduction of somatic cell nucleus into enucleated oocyte/zygote
• Cellular fusion: the fusion of ES and somatic cells (induced by viruses, chemical agents and electronic current)
• Cell explantation: the generation of pluripotent cells directly by explanting cells into appropriate culture/co-culture conditions
• Direct reprogramming: iPS
Experimental approaches for reprogramming
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TÁMOP-4.1.2-08/1/A-2009-0011Safety issues related to stem cell research/regenerative medicine• Human ES cell culture – xenogenic exposure
to mouse fibroblasts• Transmission of human viral pathogens• Error in reprogramming• Serum-derived factors
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Summary
• Stem cell research is relatively new, with substantial progress achieved and raising even more widespread hype (scientific as well as public).
• Different cells and procedures have been employed – no magic cure as yet.
• Issues to be solved: cell isolation, maintenance, reprogramming, overcoming alloreactivity and pathological differentiation.