Methods of producing pancreatic hormones
Disclosed herein are methods of producing pancreatic hormone-expressing cells by first differentiating pluripotent cells in cell culture so as to produce endodermal cells, the endodermal cells being competent to further differentiate into hormone-expressing cells capable of secreting at least one pancreatic hormone in response to a physiological signal, and then, transplanting the cultured endodermal cells into an organism, such as an organism in need of an endocrine cell therapy.
1. A method for producing hormone secreting cells in vivo, said method comprising the steps of:
a) producing human PDX-1 positive pancreatic hormone expressing cells in vitro; and
b) transplanting the human PDX-1 positive pancreatic hormone expressing cells into a mammalian subject, wherein the PDX-1 positive pancreatic hormone expressing cells secrete a hormone in vivo.
2. A method for producing hormone secreting cells, said method comprising the steps of:
a) obtaining a population of human pluripotent stem cells;
b) differentiating the population of step (a) into human definitive endoderm cells;
c) differentiating the population of human definitive endoderm cells into a population of human PDX-1 expressing pancreatic endoderm cells; and
d) transplanting the population of human PDX-1 expressing pancreatic endoderm cells into a mammal, wherein the transplanted cells of step (c) are matured into hormone producing pancreatic endocrine cells.
3. The method of claim 1 , wherein the hormone secreting cells secrete insulin.
4. The method of claim 1 , wherein the hormone secreting cells express NKX6.1.
5. The method of claim 1 , wherein the hormone secreting cells secrete glucagon.
6. The method of claim 1 , wherein the mammalian subject is a human subject.
7. The method of claim 6 , wherein the human subject has been identified as having a condition which limits the ability of the subject to produce sufficient levels of insulin in response to physiologically high blood glucose concentrations.
8. The method of claim 1 , wherein the PDX-1 positive pancreatic cells are pancreatic endoderm cells, pancreatic progenitor cells, or endocrine precursor cells.
9. The method of claim 3 , wherein insulin secretion is detected by measuring C-peptide levels.
10. The method of claim 9 , wherein C-peptide levels increase over time.
11. The method of claim 3 , wherein the insulin secreting cells are responsive to blood glucose.
12. The method of claim 3 , wherein the number of insulin secreting cells increases over time.
13. The method of claim 3 , wherein the insulin secreting cells form islet like cell clusters.
14. The method of claim 1 , wherein the transplanted PDX-1 positive pancreatic cells form pancreatic epithelium.
15. The method of claim 14 , wherein the pancreatic epithelium expresses NKX6.1.
16. The method of claim 14 , wherein the pancreatic epithelium expands over time.
17. The method of claim 2 , wherein the hormone secreting cells secrete insulin.
18. The method of claim 2 , wherein the hormone secreting cells express NKX6.1.
19. The method of claim 2 , wherein the hormone secreting cells secrete glucagon.
20. The method of claim 2 , wherein the mammal is a human subject.
21. The method of claim 20 , wherein the human subject has been identified as having a condition which limits the ability of the subject to produce sufficient levels of insulin in response to physiologically high blood glucose concentrations.
22. The method of claim 2 , wherein the PDX-1 expressing pancreatic cells are pancreatic endoderm cells, pancreatic progenitor cells, or endocrine precursor cells.
23. The method of claim 11 , wherein insulin secretion is detected by measuring C-peptide levels.
24. The method of claim 23 , wherein C-peptide levels increase over time.
25. The method of claim 17 , wherein the insulin secreting cells are responsive to blood glucose.
26. The method of claim 17 , wherein the number of insulin secreting cells increases over time.
27. The method of claim 17 , wherein the insulin secreting cells form islet like cell clusters.
28. The method of claim 2 , wherein the transplanted PDX-1 expressing pancreatic cells form pancreatic epithelium.
29. The method of claim 28 , wherein the pancreatic epithelium expresses NKX6.1.
30. The method of 28 , wherein the pancreatic epithelium expands over time.