Stem Cell-derived In Vitro Models of Receptor Mediated ... ADA Meeting . Stem Cell-derived In Vitro Models of Receptor Mediated Glucose Regulation . ... for in vitro Native Human Biology

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2014 ADA Meeting

Stem Cell-derived In Vitro Models of Receptor Mediated Glucose Regulation

David Mann, PhD CDI employee, no other relationships to disclose

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Induced Pluripotent Stem Cell Derived Tissues for in vitro Native Human Biology and Disease Modeling

iPS Cell Reprogramming

Episomal “footprint-free” method Optimized for small amounts of blood

(other tissue samples accepted)

Feeder-free, defined conditions

iPS Cell Genetic Engineering

Gene editing, deletion, or insertion Endogenous & safe harbor loci HR and nuclease technologies

iPS Cell Differentiation

Catalog cell types (and new products)

Collect Fresh Blood

Expand CD34+ Cells

Introduce Reprogramming

Factors Pick, Expand & QC iPS Cell

Clones

Cortical Neurons Cardiomyocytes Hepatocytes Endothelial Cells

Dopaminergic Neurons Skeletal Myoblast

Cardiac Progenitor Cells

Hematopoietic Progenitor Cells

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iPSCs in Modeling Native Human Biology

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iCell Hepatocytes Basic Characterization

Liver Secreted Transport Protein

Albumin PAS

Glycogen Storage

BODIPY

Lipid Storage Morphology

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iCell Hepatocytes Insulin Receptor Signaling

Modulation of pAKT (Ser473) Signal

EC50 = 2 nM

Glucose Detection

EC50 = 4 nM EC50 = 254 nM

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iCell Hepatocytes Glucagon Mediated Receptor Signaling

EC50 = 43 nM Glucagon EC50 = 2 nM

Glucose Detection cAMP Assay

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Identity Purity

iCell Skeletal Myoblasts Basic Characterization

Hoechst Troponin T Overlay

Formation of Multinucleated Myotubes

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iCell Skeletal Myoblasts Insulin Receptor Signaling (pAKT)

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iPSCs in Disease Modeling

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iCell Cardiomyocytes Induction of Diabetic Cardiomyopathy

Lipid accumulation Oxidative Stress Structural Alterations

- + - + - +

Normal Media

Diabetic Media

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iCell Cardiomyocytes Development of Diabetic Media Conditions

Beat Rate Irregularity BNP Expression (Cardiac Stress)

0 24 h 48 h 72 h 96 h0

10

20

30

40

50

60

70

Bea

t Rat

e (1

/min

)

0 24 h 48 h 72 h 96 h0

10

20

30

40

50

60

70

Bea

t Rat

e (1

/min

)

Hypertrophy Markers

Normal Media

Diabetic Media

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Generation of iPSC-Derived Diabetic Patient-Specific CMs

Control CM Type 2 Diabetic Patient CMs

Slow CVD Onset (SP)

Rapid CVD Onset (FP)

Normal (C)

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iPSC-Derived Diabetic Patient-Specific CMs Recapitulating Disease Phenotypes

C SP FP C SP FP

Structural Alterations (Normal Media)

0 24 48 72 96 120 144 1680

2

4

6

8

10

Control

01266

01267

iCPM MM DM

Time (h)

Cel

l Ind

ex (C

I)

Phenotypic Differences (Normal Media)

Slow CVD Onset (SP)

Rapid CVD Onset (FP)

Normal (C)

Proliferative Capacity

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The Future: Building Disease and Diversity Panels iPS Cell Manufacture & Banking

CIRM Human iPS Cell Initiative 3000 donors (healthy & disease phenotypes) Disease categories: epilepsy, autism, cerebral palsy, cardiomyopathy, Alzheimer’s

disease, eye diseases, hepatitis (HCV), non-alcoholic steatohepatitis (NASH), pulmonary fibrosis

Derived from peripheral blood (preferred) or skin fibroblasts Episomal “footprint-free” method Coriell Partnership for cell banking & distribution – publicly available Joint facility located within the Buck Institute, Novato, CA

CDI Interested in Partnering to Obtaining Relevant Diabetic Patient Samples MODY Genotypes Akt/Pathway Mutations Other Relevant Genotypes

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Save the Date! 3rd Annual CDI User Group Meeting

Visit us at ADA Booth 2018 And

3rd Annual CDI User Group Meeting Date: August 11-13, 2014

Location: Boston