1. Stem cells to make β cells and
recreate human Type 1 diabetes
JDRF Spring Research Briefing
Douglas Melton
Dept of Stem Cell and Regenerative
Biology
Harvard Stem Cell Institute
Howard Hughes Medical Institute

2. Pancreatic islet
insulin
glucagon
pancreatic
polypeptide
ghrelin
somatostatin

3. Pancreatic islet
insulin
glucagon
pancreatic
polypeptide
ghrelin
somatostatin

4. Diabetic dog rescued by
injection of islet extract
Best
Banting
1921

5. Type I diabetes
1. Loss of beta cells – insulin dependence
(relevance for some Type II diabetics)
Make more β cells
2. Autoimmune attack of β cells
Analyze the development of the immune
attack to induce tolerance

6. Human islets for transplantation
Photo: Dr. Gordon Weir, Joslin Diabetes Center

7. How to make pancreatic β cells
1. Directed differentiation of embryonic
stem or iPS cells
2. Direct reprogramming
3. Control β cell replication

8. 2 ways to make more to β cells
Directed differentiation β cells
ES / iPS cell
Growth factors,
Definitive endoderm Co-culture
Sox 17 Foregut
FoxA2 Pancreas
Pdx1
Endocrine
Chemical screening
Ngn3
β cell

9. High content chemical screens
on hES and mES cells
Starting ES
population 5K Chemical DMSO control Hit compound
library 4 days Sox17
O’night incubation staining
incubation
HTS image-based
analysis
Lee Rubin
HSCI

10. 2 compounds induce definitive endoderm
(IDEs)

11. IDE compounds efficiently induce definitive
endoderm
mouse > 70%
human >50%

12. 2 ways to make more to β cells
Directed differentiation β cells
ES / iPS cell
Definitive endoderm
Foregut
Compound 1
(IDEs, 80%) Pancreas
Compound 2 Endocrine
( Indolactam V, 45%)
β cell
Borowiak et al, Cell Stem Cell, 2009; Chen et al, Nat Chem Biol, 2009

13. Differentiation

14. cloning / iPS derivation

15. Direct reprogramming?

16. Pancreatic tissues
islet
vessel
duct
acini

17. Expression of 9 trans factors induces
extra islet ins+ cells

18. Reprogrammed cells express genes essential
for β cell function

19. New insulin+ cells closely resemble β cells

20. Reprogrammed cells produce and
secrete insulin

21. Type I diabetes
1. Loss of beta cells – insulin dependence
Make more β cells
2. Autoimmune attack of β cells
Analyze the development of the immune
attack to induce tolerance

22. Stem Cells in Regenerative Medicine

23. Key Components for Type I
Diabetes

24. Diabetic patient specific induced Pluripotent
Stem (DiPS) cells
OCT4
SOX2
KLF4
2 days 4-6 weeks
Biopsy DiPS
Patient Fibroblast DiPS (10x) DiPS (40x)

25. Directed differentiation β cells
2 ways to make more to thymic
epithelial cells
ES / iPS cell
Definitive endoderm
Pharyngeal foregut
Pax 9 Thymic epithelial progenitor
FoxN1
Thymic epithelial cell

26. Recapitulating human Type 1 diabetes in a
humanized mouse
DiPS
Thymic Epithelial Cells Blood Cells Pancreatic Beta Cells
George Daley
NOD, SCID, IL2-rγ
Dale Greiner
Lenny Schultz
• Function of cells population
• Disease onset and progression
• Treatments or cure

27. Patient’s perspective: Who will cure my
disease?
Pharma Hospital
JDRF
Insurance Academic
Co. (Univ)

28. Harvard Stem Cell Institute
Dept of Stem Cell and Regenerative
Biology
Howard Hughes Medical Institute
Malgorzata Borowiak Yuval Dor
Shuibing Chen Dale Greiner
Adriana Tajonar Lenny Shultz
Rene Maehr Lee Rubin
Saranya Purushothaman Stuart Schreiber
Joe Zhou George Daley
Derrick Rossi
Marc Vidal