From Bench to Bedside: Research and Clinical Applications of Induced Pluripotent Stem Cells

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

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CVC

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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