METHODS OF PRODUCING PANCREATIC HORMONES

• US 20090004152A1
• Filed: 07/05/2007
• Published: 01/01/2009
• Est. Priority Date: 03/02/2006
• Status: Active Grant

• Current Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Original Assignee: CyThera, Inc. (Vertex Pharmaceuticals Incorporated)
• Inventors: Martinson, Laura, Kroon, Evert, Baetge, Emmanuel Edward, D'Amour, Kevin
• Granted Patent: US 7,695,965 B2
• Time in Patent Office: Days
• US Class Current: 424/93.700
• 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 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) and their progeny 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 the agent is an activin selected from the group consisting of activin A, activin AB, activin B, and combinations thereof.

3. The method of claim 2, wherein the activin is activin A.

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

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

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

6. 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) transplanting the hES-derived cells of step (a) and their progeny into a mammalian subject; and

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

7. The method of claim 6, wherein the agent is an activin selected from the group consisting of activin A, activin AB, activin B, and combinations thereof.

8. The method of claim 7, wherein the activin is activin A.

9. The method of claim 6, wherein the medium lacks nicotinamide.

10. The method of claim 6 further comprising contacting the hES cells with Wnt3A.

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

(a) contacting human embryonic stem (hES) cells in vitro with a medium comprising a first 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) and their progeny into a mammalian subject; and

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

12. The method of claim 11, wherein the first agent is an activin.

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

14. The method of claim 13, wherein the activin is activin A.

15. The method of claim 11, wherein the TGF-beta family receptor is the activin receptor-like kinase (ALK) receptor.

16. The method of claim 11, wherein the second agent is SB-431542.

17. The method of claim 11, wherein the first and second medium lack nicotinamide.

19. The method of claim 11, wherein the mammalian subject is a human subject.

20. The method of claim 19, 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.

21. An in vivo insulin secreting cell derived from an in vitro human embryonic stem cell (hESC) or progeny thereof.

22. The insulin secreting cell of claim 22, wherein the in vitro hESC-derived cell or progeny thereof do not form a teratoma in vivo.