Methods of producing pancreatic hormones

• US 9,980,986 B2
• Filed: 03/26/2015
• Issued: 05/29/2018
• Est. Priority Date: 03/02/2006
• Status: Active Grant

• Current Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Original Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Inventors: Martinson, Laura, Kroon, Evert, D'Amour, Kevin, Baetge, Emmanuel Edward
• Primary Examiner(s): Ton, Thaian N
• Assistant Examiner(s): Montanari, David A.
• Application Number:
  US14/670,249
• Publication Number:
  US 20150265657A1
• Time in Patent Office: 1,160 Days
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 38/26 Glucagons
• CPC Class Codes: A61K 38/28 Insulins
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/10 Growth factors
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/113 Acidic fibroblast growth factor (aFGF, FGF-1)
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/119 Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
• CPC Class Codes: C12N 2501/12 Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/33 Insulin together with transferrin C12N2500/25; Insulin-like growth factors C12N2501/105
• CPC Class Codes: C12N 2501/385 of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
• CPC Class Codes: C12N 2501/41 Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2501/415 Wnt; Frizzeled
• CPC Class Codes: C12N 2501/42 Notch; Delta; Jagged; Serrate
• CPC Class Codes: C12N 2501/58 Adhesion molecules, e.g. ICAM, VCAM, CD18 (ligand), CD11 (ligand), CD49 (ligand)
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0678 : Stem cells; Progenitor cells; Precursor cells

Abstract

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.

Claims

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.

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.

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.

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.

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.

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.