Maintenance of islet cells

• US 20030228287A1
• Filed: 10/18/2002
• Published: 12/11/2003
• Est. Priority Date: 06/07/2002
• Status: Abandoned Application

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Beattie, Gillian M., Hayek, Alberto, Montgomery, Anthony
• Application Number:
  US10/274,529
• Publication Number:
  US 20030228287A1
• Time in Patent Office: Days
• US Class Current: 424/93.7
• 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 2533/56 Fibrin; Thrombin
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0677 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells

Abstract

The present invention provides a method of culturing cells on a matrix. The present invention includes islet cells cultured in a 3-D configuration in the presence of a fibrin matrix support. Induction of β-cell proliferation with HGF/SF is also provided.

Claims

1. A method of culturing cells, the method comprising:

culturing the cells on a matrix, wherein the matrix comprises an integrin ligand, wherein the integrin ligand is selected from the group comprising an α

5β

1 integrin ligand and an α

vβ

1 integrin ligand.

2. The method according to claim 1, wherein the integrin ligand is an α

5β

1 integrin ligand.

3. The method according to claim 1, wherein the integrin ligand is an α

{overscore (ω

)}β

1 integrin ligand

4. The method according to claim 1, wherein the matrix further comprises a synthetic mesh.

5. The method according to claim 1, wherein the integrin ligand is fibrin.

6. The method according to claim 1, wherein the integrin ligand is fibronectin.

7. The method according to claim 1, wherein the integrin ligand is vitronectin.

8. The method according to claim 1, wherein the culturing step comprises mixing fibrinogen and thrombin, thereby forming a matrix comprising fibrin.

9. The method according to claim 1, wherein the cells are islet cells.

10. The method according to claim 9, wherein the cells are human islet cells.

11. The method according to claim 1, wherein the method further comprises contacting the cells with hepatocyte growth factor.

12. The method according to claim 11, wherein the culturing step further comprises contacting the cells with:

kaposis fibroblast growth factor; and

nicotinamide.

13. The method according to claim 12, wherein the human hepatocyte growth factor is present at 20 ng/ml;

the kaposis fibroblast growth factor is present at 1 ng/ml; and

the nicotinamide is present at 10 mM.

14. The method according to claim 1, whereby dc-differentiation of the cells is prevented.

15. The method according to claim 1, whereby senescence of the cells is prevented.

16. A method of cell transplantation, the method comprising the steps of:

culturing cells on a matrix, wherein the matrix comprises an integrin ligand, wherein the integrin ligand is selected from the group comprising an α

5β

1 integrin ligand and an α

vβ

1 integrin ligand; and

administering the cells to a mammal.

17. The method according to claim 16, wherein the integrin ligand is a α

5β

1 integrin ligand.

18. The method according to claim 16, wherein the integrin ligand is a α

vβ

1 integrin ligand.

19. The method according to claim 16, wherein the cells are administered to a diabetic animal.

20. The method according to claim 16, wherein the cells are islet cells.

21. The method according to claim 16, wherein the matrix further comprises a synthetic mesh.

22. The method according to claim 16, wherein the integrin ligand is fibrin.

23. The method according to claim 16, wherein the integrin ligand is fibronectin.

24. The method according to claim 16, wherein the integrin ligand is vitronectin.

25. The method according to claim 16, wherein the culturing step comprises mixing fibrinogen and thrombin, thereby forming a matrix comprising fibrin.

26. The method according to claim 16, wherein the culturing step comprises contacting the cells with hepatocyte growth factor.

27. The method according to claim 26, wherein the culturing step further comprises contacting the cells with:

kaposis fibroblast growth factor; and

nicotinamide.

28. The method according to claim 27, wherein the human hepatocyte growth factor is present at 20 ng/ml;

the kaposis fibroblast growth factor is present at 1 ng/ml; and

the nicotinamide is present at 10 mM.

29. The method according to claim 16, further comprising cleaving the matrix with a protease before the step of administering.

30. The method according to claim 29, wherein the protease is streptokinase.

31. The method according to claim 16, wherein the administration is by implantation under a kidney capsule of the mammal.

32. The method according to claim 16, wherein the administration is subcutaneous.

33. The method according to claim 16, wherein the administration is intravenous.

34. The method according to claim 16, wherein the administration is via a liver portal vein.

35. The method according to claim 16, wherein the mammal is a human.

36. The method according to claim 16, wherein the islet cells are human islet cells.

37. The method according to claim 16, wherein the islet cells are autologous.

38. The method according to claim 16, wherein the islet cells are heterologous.

39. A composition comprising isolated cells on a fibrin matrix.

40. The method according to claim 39, wherein the cells are islet cells.

41. The method according to claim 40, wherein the cells are human islet cells.

42. The composition according to claim 39, wherein the matrix is formed by mixing fibrinogen and thrombin.

43. The composition according to claim 39, further comprising hepatocyte growth factor.

44. The composition according to claim 43, further comprising:

kaposis fibroblast growth factor; and

nicotinamide.

45. The composition according to claim 44, wherein the human hepatocyte growth factor is 20 ng/ml;

the kaposis fibroblast growth factor is 1 ng/ml; and

the nicotinamide is 10 mM.