Post on 16-Jul-2015
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From Bench to Bedside:
Research and Clinical
Applications of Induced
Pluripotent Stem Cells
Created and Narrated by
Theresa Gold
Hematopoietic Stem Cells
Neural Stem Cells
Mesenchymal Stem Cells
Blood Cells Cells of the Nervous System
Connective Tissue, Bones, Cartilage.
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Embryonic Stem Cells Adult Stem Cells
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Trophoblast
Inner Cell Mass
The Blastocyst
Pancreatic β-Cells
Erythrocytes
Dopaminergic Neurons
Purified Embryonic Stem Cell Culture
Hematopoietic Stem Cells
Multipotent Stem Cells
Lymphoid Progenitor Cell
Myeloid Progenitor Cell
Erythrocytes
Neutrophil
Basophil
Adult Stem Cells
Donor HLA
Recipient Antibodies to HLA
Pancreatic β-Cells
Dopaminergic Neurons
Erythrocytes
Cardiac Cells
Parkinson’s Disease
Cardiac Failure
Diabetes Mellitus
Sickle-cell Anemia
Oct3/4
Sox2
C-Myc
Klf4
Expression of Oct3/4, Sox2, c-Myc, Klf4
Somatic Cell Donor
Isolation of fibroblasts
Fibroblast Culture
Viral transduction of pluripotency factors:
• Oct3/4• Sox2• C-Myc• Klf4
iPs Cells
Induced pluripotent stem cells are morphologically indistinguishable from embryonic stem cells.
Embryonic Stem Cells
Induced Pluripotent Stem Cells
Mouse Embryonic Fibroblasts
Microarray analysis of iPS cells revealed slight differences in gene expression profiles of iPS and embryonic stem cells.
Chromatin immunoprecipitation assay revealed differences in methylation and acetylation patterns of the promoter regions of pluripotency and development associated genes between iPS and ES cells.
Tissues from all three germ layers present in teratomas derived from iPS cells.
iPS cells were microinjected into
blastocysts. Embryos were analyzed with a
fluorescence microscope
The chimeric embryo was sectioned and
stained with anti-GFP antibody (brown). Cells
were counterstained with eosin (blue).Chimeric embryos
contained cell from all three germ layers.
Contribution of iPS cells to mouse embryonic development.
Biopsy
ReprogrammingCardiac Cells
Hepatocytes
Neuron
Cardiac Cells
Hepatocytes
Neuron
Genetic Repairof iPS Cells
Differentiationof iPS Cells
Drug screening
and disease modeling
Differentiation of iPS Cells
Up-regulation of adjacent genes
Retroviral Transduction
Plasmid Transfection
Direct Delivery of Reprogramming Proteins
Isolation of fibroblasts
Fibroblast Culture
Addition of pluripotency factors:
• Oct3/4• Sox2• C-Myc• Klf4
iPs Cells
Differentiation into Pancreatic β-Cells
Diabetes Mellitus Patient
Transplantation of Pancreatic β-Cells
Patient derived Induced Pluripotent
Stem Cells
Patient HLA
Patient Antibodies-do not react with own HLA
CV C
V
CVC
VCV
CV
CV
CV
Parkinson’s Disease
Inside the BrainDopaminergic neurons within the substantia
nigra degenerate.
Dopaminergic Neurons
SymptomsCognitive: loss of
executive functions.Physical: tremors,
stiffness, and slowed movement
Fibroblast Culture
Retroviral transduction of
pluripotency factors
• Oct3/4• Sox2• C-Myc• Klf4
iPS Cells
Neuronal Differentiation
• Sonic Hedgehog• Fibroblast Growth
Factor 8
Dopaminergic Neurons
Transplantation of Dopaminergic Neurons
iPS cell treated rats display improved behavior and motor function
Isolation of Fibroblasts
Rat Model of Parkinson’s Disease
Isolation of fibroblasts
Parkinson’s Disease Patient
Fibroblast Culture
Reprogramming
iPS Cell Culture
Differentiation
Dopaminergic Neurons
Potential Applications of Induced Pluripotent Stem Cells
StrokeTraumatic injury
Alzheimer's DiseaseParkinson’s Disease
Bone marrow transplantation
Blindness
Deafness
Myocardial infarction
Muscular dystrophy
Diabetes mellitusSpinal cord injury