METHODS OF PRODUCING PANCREATIC HORMONES

• US 20080267926A1
• Filed: 07/02/2008
• Published: 10/30/2008
• Est. Priority Date: 07/26/2006
• Status: Active Grant

• Current Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Original Assignee: CyThera, Inc. (Vertex Pharmaceuticals Incorporated)
• Inventors: Martinson, Laura, Kroon, Evert, D'Amour, Kevin, Baetge, Emmanuel E.
• Granted Patent: US 7,534,608 B2
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• 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 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• 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 insulin, said method comprising the steps of:

(a) contacting human embryonic stem (hES) cells in vitro with a first medium comprising an agent that activates a TGF-β

receptor family member;

(b) culturing in vitro the hES-derived cells of step (a) in a second medium lacking the agent that activates the TGF-β

receptor family member;

(c) transplanting the hES-derived cells of step (b) into a mammalian subject; and

(d) maturing the hES-derived cells of step (c) in vivo, thereby obtaining insulin secreting cells.

2. The method of claim 1, wherein said second medium comprises an agent that inhibits the TGF-β

receptor family member.

3. The method of claim 1, wherein the agent comprises an activin selected from the group consisting of activin A, activin AB, activin B, and combinations thereof.

4. The method of claim 1, wherein hES-derived cells of step (b) comprise endocrine precursor cells.

5. The method of claim 1, wherein the first and second medium lacks nicotinamide.

6. The method of claim 1, wherein the TGF-β

family receptor is the activin receptor-like kinase (ALK) receptor.

7. The method of claim 1 further comprising contacting the hES cells with Wnt3A.

8. The method of claim 1, wherein the mammalian subject is a human subject.

9. The method of claim 8, 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.

10. The method of claim 1, wherein hES-derived cells of step (b) comprise PDX-1 positive pancreatic endoderm cells.

11. The method of claim 1, wherein the insulin secreting cell secretes insulin in response to glucose stimulation.

12. A method for producing insulin, said method comprising the steps of:

(a) contacting human embryonic stem (hES) cells in vitro with a medium comprising an agent that activates a TGF-β

receptor family member

(b) culturing in vitro the hES-derived cells of step (a) in a second medium comprising a second agent that inhibits the TGF-β

receptor family member;

(c) transplanting the hES-derived cells of step (b) into a mammalian subject; and

(d) maturing the hES-derived cells of step (c) in vivo, thereby obtaining insulin secreting cells.

13. The method of claim 12, wherein the first agent is selected from the group consisting of activin A, activin AB, activin B and combinations thereof.

14. The method of claim 12, wherein hES-derived cells of step (b) comprise at least one of the group comprising endocrine precursor cells and PDX-1 positive pancreatic endoderm cells.

15. The method of claim 12, wherein the TGF-β

family receptor is the activin receptor-like kinase (ALK) receptor.

16. The method of claim 12, wherein the second agent is SB-431542.

17. The method of claim 12 further comprising contacting the hES cells with Wnt3A.

18. The method of claim 12, wherein the mammalian subject is a human subject.

19. The method of claim 18, 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.

20. The method of claim 12, wherein the insulin secreting cell secretes insulin in response to glucose stimulation.