Compositions and methods for establishing and maintaining stem cells in an undifferentiated state

• US 20100317100A1
• Filed: 06/11/2010
• Published: 12/16/2010
• Est. Priority Date: 06/12/2009
• Status: Active Grant

• Current Assignee: University of Kansas
• Original Assignee: University of Kansas
• Inventors: Paul, Soumen, Dutta, Debasree, Schoenen, Frank John, Ray, Soma, Aube, Jeffrey
• Granted Patent: US 8,193,235 B2
• Time in Patent Office: Days
• US Class Current: 435/353
• CPC Class Codes: C07D 403/04 directly linked by a ring-member-to-ring-member bond
• CPC Class Codes: C07D 403/14 containing three or more hetero rings
• CPC Class Codes: C12N 2501/727 Kinases (EC 2.7.)
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

The present invention embraces compositions and methods for establishing and maintaining stem cells and inhibiting stem cell differentiation using a selective Protein Kinase C (PKC) inhibitor.

Claims

1. A PKC inhibitor of Formula I, II, III, IV, or V,

2. A method for maintaining a stem cell in an undifferentiated state comprising contacting an isolated stem cell with a selective Protein Kinase C inhibitor thereby maintaining the stem cell in an undifferentiated state.

3. The method of claim 2, wherein the inhibitor inhibits at least Protein Kinase C zeta.

4. The method of claim 3, wherein the inhibitor further inhibits Protein Kinase C alpha and delta.

5. The method of claim 2, wherein the inhibitor concentration is in the range of 100 nm to 5 μ

M.

6. The method of claim 2, wherein the undifferentiated state is maintained in the absence of a feeder cell or leukemia inhibitory factor.

7. The method of claim 2, wherein the stem cell is isolated from a mouse.

8. The method of claim 2, wherein the stem cell is isolated from a rat.

9. The method of claim 2, wherein the stem cell is isolated from a human.

10. The method of claim 2, wherein the stem cell is an embryonic stem cell, adult stem cell, induced pluripotent stem cell, or cancer stem cell.

11. The method of claim 2, wherein the PKC inhibitor is a PKC inhibitor of Formula I, II, III, IV, or V,

12. A method for inhibiting differentiation of a stem cell comprising contacting an isolated stem cell with a selective Protein Kinase C inhibitor thereby inhibiting differentiation of the stem cell.

13. The method of claim 12, wherein the inhibitor inhibits at least Protein Kinase C zeta.

14. The method of claim 13, wherein the inhibitor further inhibits Protein Kinase C alpha and delta.

15. The method of claim 12, wherein the inhibitor concentration is in the range of 100 nm to 5 μ

M.

16. The method of claim 12, wherein differentiation is inhibited under differentiation conditions.

17. The method of claim 12, wherein the stem cell is isolated from a mouse.

18. The method of claim 12, wherein the stem cell is isolated from a rat.

19. The method of claim 12, wherein the stem cell is isolated from a human.

20. The method of claim 12, wherein the stem cell is an embryonic stem cell, adult stem cell, induced pluripotent stem cell, or cancer stem cell.

21. The method of claim 12, wherein the PKC inhibitor is a PKC inhibitor of Formula I, II, III, IV, or V,

22. A method for establishing a pluripotent stem cell line comprising culturing a cell with a selective Protein Kinase C inhibitor thereby establishing a pluripotent stem cell line.

23. The method of claim 22, wherein the inhibitor inhibits as least Protein Kinase C zeta.

24. The method of claim 22, wherein the inhibitor further inhibits Protein Kinase C alpha and delta.

25. The method of claim 22, wherein the inhibitor concentration is in the range of 100 nm to 5 μ

M.

26. The method of claim 22, wherein the cell is isolated from a mouse.

27. The method of claim 22, wherein the cell is isolated from a rat.

28. The method of claim 22, wherein the cell is isolated from a human.

29. The method of claim 22, wherein the cell is a blastocyte or fibroblast.

30. The method of claim 22, wherein the PKC inhibitor is a PKC inhibitor of Formula I, II, III, IV, or V,