stem cells hessah alshammari. stem cells are distinguished from other cell types by two important...
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Stem cells
Hessah Alshammari
bull Stem cells are distinguished from other cell types by two important characteristics
bull They are unspecialized cells capable of renewing themselves through cell division sometimes after long periods of inactivity
bull Under certain conditions they can be induced to become tissue- or organ-specific cells with special functions eg in bone marrow stem cells regularly divide to repair and replace worn out or damaged tissues
Stem cells types
bull Embryonic stem cells
bull Adult stem cells
bull Cancer stem cells
Embryonic stem cells
bull Embryonic stem cells derived from embryo in typically four or five days old which called the blastocyst
bull The blastocyst includes three structures the trophoblast which is the layer of cells that surrounds the blastocoel a hollow cavity inside the blastocyst and the inner cell mass which is a group of cells at one end of the blastocoel that develop into the embryo proper
bull An adult stem cell is an undifferentiated cell found among differentiated cells in a tissue or organ that can renew itself and can differentiate to yield some or all of the major specialized cell types of the tissue or organ
bull The primary roles of adult stem cells in a living organism are to maintain and repair the tissue
bull In fact adult hematopoietic or blood-forming stem cells from bone marrow have been used in transplants for 40 years
Adult stem cells
Similarities and differences between embryonic and adult stem cells
Embryonic stem cells adult stem cells
Differentiation ability
Pluripotent stem cellscan become all cell types of the body
multipotent stem cellslimited to differentiating into different cell types of their tissue
culture grown easily in culture Isolation and expansion challenging
Rejection after transplantation
Do not cause rejection cause rejection
Tumour formation after injection
tumour genesis is not tumour genesis
Colony forming form embryoid bodys MSC form colony forming units NSC form neurispheres
embryoid bodys
colony forming units
Neurispheres
Adult stem cells in human body
bull Neural stem cellsbull Heamatopeotic stem cellsbullMesenchymal stem cellsbullAdipose stem cellsbullSkin stem cellsbullLiver stem cellsbullMultipotent adult precursor cells MAPC bullMammary stem cells
Mesenchymal stem cell
bullSomatic progenitors able to self-renew and produce mesenchymal derivatives such as osteroblast adipocytes and chondrocytes
bullCD29 amp CD105 is MSCs marker
Mesenchymal stem cell isolation
Isolation of bone marrow
Lysis blood cells using buffer and culture
MSC
Biomedical applications
1- Pharmacological screening
2- Orthopaedic tissue repair (bone cartilage tendon)
3- Non-orthopaedic application (heartspinal cord)
4- Gene delivery
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
bull Stem cells are distinguished from other cell types by two important characteristics
bull They are unspecialized cells capable of renewing themselves through cell division sometimes after long periods of inactivity
bull Under certain conditions they can be induced to become tissue- or organ-specific cells with special functions eg in bone marrow stem cells regularly divide to repair and replace worn out or damaged tissues
Stem cells types
bull Embryonic stem cells
bull Adult stem cells
bull Cancer stem cells
Embryonic stem cells
bull Embryonic stem cells derived from embryo in typically four or five days old which called the blastocyst
bull The blastocyst includes three structures the trophoblast which is the layer of cells that surrounds the blastocoel a hollow cavity inside the blastocyst and the inner cell mass which is a group of cells at one end of the blastocoel that develop into the embryo proper
bull An adult stem cell is an undifferentiated cell found among differentiated cells in a tissue or organ that can renew itself and can differentiate to yield some or all of the major specialized cell types of the tissue or organ
bull The primary roles of adult stem cells in a living organism are to maintain and repair the tissue
bull In fact adult hematopoietic or blood-forming stem cells from bone marrow have been used in transplants for 40 years
Adult stem cells
Similarities and differences between embryonic and adult stem cells
Embryonic stem cells adult stem cells
Differentiation ability
Pluripotent stem cellscan become all cell types of the body
multipotent stem cellslimited to differentiating into different cell types of their tissue
culture grown easily in culture Isolation and expansion challenging
Rejection after transplantation
Do not cause rejection cause rejection
Tumour formation after injection
tumour genesis is not tumour genesis
Colony forming form embryoid bodys MSC form colony forming units NSC form neurispheres
embryoid bodys
colony forming units
Neurispheres
Adult stem cells in human body
bull Neural stem cellsbull Heamatopeotic stem cellsbullMesenchymal stem cellsbullAdipose stem cellsbullSkin stem cellsbullLiver stem cellsbullMultipotent adult precursor cells MAPC bullMammary stem cells
Mesenchymal stem cell
bullSomatic progenitors able to self-renew and produce mesenchymal derivatives such as osteroblast adipocytes and chondrocytes
bullCD29 amp CD105 is MSCs marker
Mesenchymal stem cell isolation
Isolation of bone marrow
Lysis blood cells using buffer and culture
MSC
Biomedical applications
1- Pharmacological screening
2- Orthopaedic tissue repair (bone cartilage tendon)
3- Non-orthopaedic application (heartspinal cord)
4- Gene delivery
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Stem cells types
bull Embryonic stem cells
bull Adult stem cells
bull Cancer stem cells
Embryonic stem cells
bull Embryonic stem cells derived from embryo in typically four or five days old which called the blastocyst
bull The blastocyst includes three structures the trophoblast which is the layer of cells that surrounds the blastocoel a hollow cavity inside the blastocyst and the inner cell mass which is a group of cells at one end of the blastocoel that develop into the embryo proper
bull An adult stem cell is an undifferentiated cell found among differentiated cells in a tissue or organ that can renew itself and can differentiate to yield some or all of the major specialized cell types of the tissue or organ
bull The primary roles of adult stem cells in a living organism are to maintain and repair the tissue
bull In fact adult hematopoietic or blood-forming stem cells from bone marrow have been used in transplants for 40 years
Adult stem cells
Similarities and differences between embryonic and adult stem cells
Embryonic stem cells adult stem cells
Differentiation ability
Pluripotent stem cellscan become all cell types of the body
multipotent stem cellslimited to differentiating into different cell types of their tissue
culture grown easily in culture Isolation and expansion challenging
Rejection after transplantation
Do not cause rejection cause rejection
Tumour formation after injection
tumour genesis is not tumour genesis
Colony forming form embryoid bodys MSC form colony forming units NSC form neurispheres
embryoid bodys
colony forming units
Neurispheres
Adult stem cells in human body
bull Neural stem cellsbull Heamatopeotic stem cellsbullMesenchymal stem cellsbullAdipose stem cellsbullSkin stem cellsbullLiver stem cellsbullMultipotent adult precursor cells MAPC bullMammary stem cells
Mesenchymal stem cell
bullSomatic progenitors able to self-renew and produce mesenchymal derivatives such as osteroblast adipocytes and chondrocytes
bullCD29 amp CD105 is MSCs marker
Mesenchymal stem cell isolation
Isolation of bone marrow
Lysis blood cells using buffer and culture
MSC
Biomedical applications
1- Pharmacological screening
2- Orthopaedic tissue repair (bone cartilage tendon)
3- Non-orthopaedic application (heartspinal cord)
4- Gene delivery
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Embryonic stem cells
bull Embryonic stem cells derived from embryo in typically four or five days old which called the blastocyst
bull The blastocyst includes three structures the trophoblast which is the layer of cells that surrounds the blastocoel a hollow cavity inside the blastocyst and the inner cell mass which is a group of cells at one end of the blastocoel that develop into the embryo proper
bull An adult stem cell is an undifferentiated cell found among differentiated cells in a tissue or organ that can renew itself and can differentiate to yield some or all of the major specialized cell types of the tissue or organ
bull The primary roles of adult stem cells in a living organism are to maintain and repair the tissue
bull In fact adult hematopoietic or blood-forming stem cells from bone marrow have been used in transplants for 40 years
Adult stem cells
Similarities and differences between embryonic and adult stem cells
Embryonic stem cells adult stem cells
Differentiation ability
Pluripotent stem cellscan become all cell types of the body
multipotent stem cellslimited to differentiating into different cell types of their tissue
culture grown easily in culture Isolation and expansion challenging
Rejection after transplantation
Do not cause rejection cause rejection
Tumour formation after injection
tumour genesis is not tumour genesis
Colony forming form embryoid bodys MSC form colony forming units NSC form neurispheres
embryoid bodys
colony forming units
Neurispheres
Adult stem cells in human body
bull Neural stem cellsbull Heamatopeotic stem cellsbullMesenchymal stem cellsbullAdipose stem cellsbullSkin stem cellsbullLiver stem cellsbullMultipotent adult precursor cells MAPC bullMammary stem cells
Mesenchymal stem cell
bullSomatic progenitors able to self-renew and produce mesenchymal derivatives such as osteroblast adipocytes and chondrocytes
bullCD29 amp CD105 is MSCs marker
Mesenchymal stem cell isolation
Isolation of bone marrow
Lysis blood cells using buffer and culture
MSC
Biomedical applications
1- Pharmacological screening
2- Orthopaedic tissue repair (bone cartilage tendon)
3- Non-orthopaedic application (heartspinal cord)
4- Gene delivery
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
bull An adult stem cell is an undifferentiated cell found among differentiated cells in a tissue or organ that can renew itself and can differentiate to yield some or all of the major specialized cell types of the tissue or organ
bull The primary roles of adult stem cells in a living organism are to maintain and repair the tissue
bull In fact adult hematopoietic or blood-forming stem cells from bone marrow have been used in transplants for 40 years
Adult stem cells
Similarities and differences between embryonic and adult stem cells
Embryonic stem cells adult stem cells
Differentiation ability
Pluripotent stem cellscan become all cell types of the body
multipotent stem cellslimited to differentiating into different cell types of their tissue
culture grown easily in culture Isolation and expansion challenging
Rejection after transplantation
Do not cause rejection cause rejection
Tumour formation after injection
tumour genesis is not tumour genesis
Colony forming form embryoid bodys MSC form colony forming units NSC form neurispheres
embryoid bodys
colony forming units
Neurispheres
Adult stem cells in human body
bull Neural stem cellsbull Heamatopeotic stem cellsbullMesenchymal stem cellsbullAdipose stem cellsbullSkin stem cellsbullLiver stem cellsbullMultipotent adult precursor cells MAPC bullMammary stem cells
Mesenchymal stem cell
bullSomatic progenitors able to self-renew and produce mesenchymal derivatives such as osteroblast adipocytes and chondrocytes
bullCD29 amp CD105 is MSCs marker
Mesenchymal stem cell isolation
Isolation of bone marrow
Lysis blood cells using buffer and culture
MSC
Biomedical applications
1- Pharmacological screening
2- Orthopaedic tissue repair (bone cartilage tendon)
3- Non-orthopaedic application (heartspinal cord)
4- Gene delivery
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Similarities and differences between embryonic and adult stem cells
Embryonic stem cells adult stem cells
Differentiation ability
Pluripotent stem cellscan become all cell types of the body
multipotent stem cellslimited to differentiating into different cell types of their tissue
culture grown easily in culture Isolation and expansion challenging
Rejection after transplantation
Do not cause rejection cause rejection
Tumour formation after injection
tumour genesis is not tumour genesis
Colony forming form embryoid bodys MSC form colony forming units NSC form neurispheres
embryoid bodys
colony forming units
Neurispheres
Adult stem cells in human body
bull Neural stem cellsbull Heamatopeotic stem cellsbullMesenchymal stem cellsbullAdipose stem cellsbullSkin stem cellsbullLiver stem cellsbullMultipotent adult precursor cells MAPC bullMammary stem cells
Mesenchymal stem cell
bullSomatic progenitors able to self-renew and produce mesenchymal derivatives such as osteroblast adipocytes and chondrocytes
bullCD29 amp CD105 is MSCs marker
Mesenchymal stem cell isolation
Isolation of bone marrow
Lysis blood cells using buffer and culture
MSC
Biomedical applications
1- Pharmacological screening
2- Orthopaedic tissue repair (bone cartilage tendon)
3- Non-orthopaedic application (heartspinal cord)
4- Gene delivery
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
embryoid bodys
colony forming units
Neurispheres
Adult stem cells in human body
bull Neural stem cellsbull Heamatopeotic stem cellsbullMesenchymal stem cellsbullAdipose stem cellsbullSkin stem cellsbullLiver stem cellsbullMultipotent adult precursor cells MAPC bullMammary stem cells
Mesenchymal stem cell
bullSomatic progenitors able to self-renew and produce mesenchymal derivatives such as osteroblast adipocytes and chondrocytes
bullCD29 amp CD105 is MSCs marker
Mesenchymal stem cell isolation
Isolation of bone marrow
Lysis blood cells using buffer and culture
MSC
Biomedical applications
1- Pharmacological screening
2- Orthopaedic tissue repair (bone cartilage tendon)
3- Non-orthopaedic application (heartspinal cord)
4- Gene delivery
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Adult stem cells in human body
bull Neural stem cellsbull Heamatopeotic stem cellsbullMesenchymal stem cellsbullAdipose stem cellsbullSkin stem cellsbullLiver stem cellsbullMultipotent adult precursor cells MAPC bullMammary stem cells
Mesenchymal stem cell
bullSomatic progenitors able to self-renew and produce mesenchymal derivatives such as osteroblast adipocytes and chondrocytes
bullCD29 amp CD105 is MSCs marker
Mesenchymal stem cell isolation
Isolation of bone marrow
Lysis blood cells using buffer and culture
MSC
Biomedical applications
1- Pharmacological screening
2- Orthopaedic tissue repair (bone cartilage tendon)
3- Non-orthopaedic application (heartspinal cord)
4- Gene delivery
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Mesenchymal stem cell
bullSomatic progenitors able to self-renew and produce mesenchymal derivatives such as osteroblast adipocytes and chondrocytes
bullCD29 amp CD105 is MSCs marker
Mesenchymal stem cell isolation
Isolation of bone marrow
Lysis blood cells using buffer and culture
MSC
Biomedical applications
1- Pharmacological screening
2- Orthopaedic tissue repair (bone cartilage tendon)
3- Non-orthopaedic application (heartspinal cord)
4- Gene delivery
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Mesenchymal stem cell isolation
Isolation of bone marrow
Lysis blood cells using buffer and culture
MSC
Biomedical applications
1- Pharmacological screening
2- Orthopaedic tissue repair (bone cartilage tendon)
3- Non-orthopaedic application (heartspinal cord)
4- Gene delivery
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Biomedical applications
1- Pharmacological screening
2- Orthopaedic tissue repair (bone cartilage tendon)
3- Non-orthopaedic application (heartspinal cord)
4- Gene delivery
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Pharmacological screening
Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)
Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Orthopaedic tissue repair (bone cartilage tendon)
The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON
MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND
Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton
Radiographs of a segmental defect that had been treated with a ceramic cylinder that
had not been loaded with MSCs(A) and loaded with
MSC (B)
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Non-orthopaedic application (heartspinal cord)
Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction
Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare
MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)
Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Adipose stem cells
Adipose-derived stem cells was reported first time in 2002 by Hedrick et al
CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC
Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Isolation of adipose stem cells
Lipoaspirate
collagenease digest
washdensity gradient centrifugation
plating
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Morphology and multilineage differentiation potential of ASCs
Primary ASCs exhibited a
fibroblast-like morphology as observed under a phase contrast
microscope
chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition
osteogenic differentiation was revealed
by ALP staining
adipogenic differentiatio
n was confirmed by positive Oil
Red O staining
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Adipose stem cells application
bullPlastic amp cosmetic application eg breast reconstruction
bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Adipose stem cells applications
Adipose-derived adult stromal cells heal critical-size mouse calvarial defects
Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Neural stem cells in brain
Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus
neural stem cells markers NestinSox1 Sox2 and Sox9
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Neural stem cells
bull Precursors for neuronal and glial lineages
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Isolation and culture of Neural stem cells
Brain digestiondetermine the
location of NSC in brain
Tissue disaggregation
by using enzyme eg accumax
Culture NSCIn serum-free medium with
EGF and bFGF
NSC form neurosphere in culture and has Sox1 amp Sox2 marker
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Neural stem cells application
bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes
Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea
bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week
Phenotype is significantly improved
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Limitations of neural stem cells application
bull Ethics of collection
bull Availability of healthy tissue
bull Viability of isolated cells
bull immunocompatibility
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Skin stem cells
Stem cells have been found in three regions of the epidermis
bull the bulge region of the follicle
bull The sebaceous gland
bull The interfollicular epidermis
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Skin stem cells properties
bull Have a high colony-forming ability in vitro
bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Skin stem cells isolation
Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
Skin stem cells applications
bull Burns and skin damage
bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010
bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis
bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt
bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature
bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged
bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
bull Genetic conditions ( epidermolysis bullosa)
Mutation in laminin 5 gene (basement membrane component)
Epidermolysis Bullosa
Extensive blistering of the skin generating infected lesions
Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
bull Genetic conditions ( epidermolysis bullosa)
Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)
They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-
- Stem cells Hessah Alshammari
- Slide 2
- Stem cells types
- Embryonic stem cells
- Adult stem cells
- Similarities and differences between embryonic and adult stem cells
- Slide 7
- Adult stem cells in human body
- Mesenchymal stem cell
- Mesenchymal stem cell isolation
- Biomedical applications
- Pharmacological screening
- Orthopaedic tissue repair (bone cartilage tendon)
- Non-orthopaedic application (heartspinal cord)
- Adipose stem cells
- Isolation of adipose stem cells
- Morphology and multilineage differentiation potential of ASCs
- Adipose stem cells application
- Adipose stem cells applications
- Neural stem cells in brain
- Neural stem cells
- Isolation and culture of Neural stem cells
- Neural stem cells application
- Limitations of neural stem cells application
- Skin stem cells
- Skin stem cells properties
- Skin stem cells isolation
- Skin stem cells applications
- Slide 29
- Slide 30
- Slide 31
-