COMPOSITION FOR CULTURING PLURIPOTENT STEM CELLS AND USE THEREOF

• US 20120164729A1
• Filed: 12/20/2011
• Published: 06/28/2012
• Est. Priority Date: 12/24/2010
• Status: Abandoned Application

• Current Assignee: National University Corporation Chiba University
• Original Assignee: National University Corporation Chiba University
• Inventors: TOMIZAWA, MINORU
• Application Number:
  US13/331,311
• Publication Number:
  US 20120164729A1
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2501/727 Kinases (EC 2.7.)
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

A means for feeder-free culture which can sufficiently maintain the undifferentiation potency of pluripotent stem cells such as iPS cells even without using heterologously derived cells or proteins, is provided.

Disclosed are a composition for pluripotent stem cell culture containing activin (preferably, activin A); a medium for pluripotent stem cell culture containing the composition; a method for culturing pluripotent stem cells for proliferating or establishing pluripotent stem cells such as iPS cells (preferably, mammalian iPS cells, and particularly preferably, human iPS cells) while maintaining an undifferentiated condition of the pluripotent stem cells, the method including performing the culture in the presence of activin A; and a method for preparing a pluripotent stem cell clonal population in an undifferentiated condition, the method including culturing undifferentiated pluripotent stem cells such as iPS cells (preferably, mammalian iPS cells, and particularly preferably, human iPS cells) in the presence of activin.

Claims

1. A composition for pluripotent stem cell culture, comprising activin.

2. The composition according to claim 1, wherein the composition is a medium supplement.

3. The composition according to claim 1, wherein the composition is intended for proliferating pluripotent stem cells while maintaining an undifferentiated condition of the pluripotent stem cells.

4. The composition according to claim 1, wherein the activin is activin A.

5. A medium for pluripotent stem cell culture, comprising the composition according to claim 1.

6. The medium according to claim 5, comprising activin at a concentration of 3 to 30 ng/mL.

7. The medium according to claim 5, wherein the medium is free of supporting cells and/or serum.

8. The medium according to claim 7, wherein the medium is free of supporting cells and serum.

9. The medium according to claim 5, wherein the medium is a cell culture minimum medium.

10. A method for culturing pluripotent stem cells for proliferating or establishing pluripotent stem cells while maintaining an undifferentiated condition of the pluripotent stem cells, the method comprising performing the culture in the presence of activin.

11. The method according to claim 10, wherein the culture is carried out in a medium containing activin.

12. The method according to claim 10, wherein the pluripotent stem cells are induced pluripotent stem cells.

13. The method according to claim 10, wherein the pluripotent stem cells are cells derived from a mammal.

14. The method according to claim 13, wherein the pluripotent stem cells are human-derived cells.

15. A method for preparing a pluripotent stem cell clonal population in an undifferentiated condition, the method comprising culturing undifferentiated pluripotent stem cells in the presence of activin.

17. The method according to claim 15, wherein the culture is carried out in a medium containing activin.

18. The method according to claim 15, wherein a single pluripotent stem cell is cultured, and thereby a clonal population of the cells is obtained.

19. The method according to claim 15, wherein pluripotent stem cells that are under lower density seeding conditions than those inducing undifferentiated proliferation of the pluripotent stem cells by the interaction between neighboring pluripotent stem cells, are cultured in a medium containing activin and free of supporting cells and/or serums, and thereby a clonal population of the cells is obtained.

20. The method according to claim 15, wherein a single pluripotent stem cell is cultured in a medium containing activin and free of supporting cells and/or serums, and a clonal population of the cell is obtained.

21. The method according to claim 15, wherein the pluripotent stem cells are induced pluripotent stem cells.

22. The method according to claim 15, wherein the pluripotent stem cells are cells derived from a mammal.

23. The method according to claim 22, wherein the pluripotent stem cells are human-derived cells.

16. A method for preparing a pluripotent stem cell clonal population in an undifferentiated condition, the method comprising isolating pluripotent stem cells in an undifferentiated condition from a living organism, and culturing undifferentiated pluripotent stem cells in the presence of activin.

24-26. -26. (canceled)