Method for generating pancreatic hormone-producing cells

Patent metadata

Field	Value
Publication number	US-9796962-B2
Application number	US-201414909690-A
Country	US
Kind code	B2
Filing date	Aug 6, 2014
Priority date	Aug 7, 2013
Publication date	Oct 24, 2017
Grant date	Oct 24, 2017

Abstract

Provided is a method for inducing pancreatic hormone-producing cells from pancreatic progenitor cells efficiently. The method comprises a step of culturing the cells in a culture medium comprising sodium cromoglicate.

First claim

The invention claimed is:

1. A method for generating pancreatic hormone-producing cells, which comprises culturing pancreas progenitor cells in a medium comprising sodium cromoglicate.

2. The method of claim 1 , wherein the medium further comprises at least one agent selected from the group consisting of:
(a) an cAMP analog;
(b) nicotinamide;
(c) a steroid; and
(d) a TGFβ inhibitor.

3. The method of claim 1 , wherein the medium further comprises:
(a) at least one agent selected from the group consisting of an adenylate cyclase activator, a cAMP phosphodiesterase inhibitor and an cAMP analog;
(b) nicotinamide;
(c) a steroid; and
(d) a TGFβ inhibitor.

4. The method of claim 2 , wherein the cAMP analog is forskolin.

5. The method of claim 2 , wherein the steroid is dexamethasone.

6. The method of claim 2 , wherein the TGFβ inhibitor is 2-(3-(6-Methylpyridin-2-yl)-1H-pyrazol-4-yl)-1,5-naphthyridine.

7. The method of claim 1 , wherein the pancreatic progenitor cells are the cells derived from a method comprising the following steps:
(1) culturing pluripotent stem cells in a medium comprising an activator of activin receptor-like kinase-4,7 and a GSK3 inhibitor, and
(2) culturing the cells obtained in step (1) in a medium comprising (a) a retinoic acid receptor agonist, (b) a BMP inhibitor and (c) a TGFβ inhibitor.

8. The method of claim 7 , wherein the activator of activin receptor-like kinase-4,7 is activin.

9. The method of claim 7 , wherein the GSK3 inhibitor is CHIR99021.

10. The method of claim 7 , wherein the TGFβ inhibitor used in step (2) is SB431542.

11. The method of claim 7 , wherein the BMP inhibitor is dorsomorphin.

12. The method of claim 1 , wherein the pancreatic hormone-producing cells are selected from the group consisting of insulin producing cells, glucagon producing cells, somatostatin producing cells and pancreatic polypeptide producing cells.

13. The method of claim 12 , wherein the pancreatic hormone-producing cells are insulin producing cells and/or glucagon producing cells.

14. The method of claim 1 , wherein the pancreatic progenitor cells are human cells.

15. The method of claim 1 , wherein the medium further comprises at least one agent selected from the group consisting of:
(a) an adenylate cyclase activator;
(b) nicotinamide;
(c) a steroid; and
(d) a TGFβ inhibitor.

16. The method of claim 15 , wherein the steroid is dexamethasone.

17. The method of claim 15 , wherein the TGFβ inhibitor is 2-(3-(6-Methylpyridin-2-yl)-1H-pyrazol-4-yl)-1,5-naphthyridine.

18. The method of claim 1 , wherein the medium further comprises at least one agent selected from the group consisting of:
(a) a cAMP phosphodiesterase inhibitor;
(b) nicotinamide;
(c) a steroid; and
(d) a TGFβ inhibitor.

19. The method of claim 18 , wherein the steroid is dexamethasone.

20. The method of claim 18 , wherein the TGFβ inhibitor is 2-(3-(6-Methylpyridin-2-yl)-1H-pyrazol-4-yl)-1,5-naphthyridine.

Assignees

• Univ Kyoto

Inventors

• Osafune Kenji
• Inagaki Nobuya
• Toyoda Taro
• Kondo Yasushi

Classifications

• C12N2501/01

  Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin · CPC title

• C12N2501/82

  Histamine · CPC title

• C12N2501/385

  of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR] · CPC title

• C12N5/0676 Primary

  Pancreatic cells · CPC title

• C12N2501/15

  Transforming growth factor beta (TGF-β) · CPC title