Cell populations which co-express CD49c and CD90

• US 8,486,696 B2
• Filed: 02/10/2005
• Issued: 07/16/2013
• Est. Priority Date: 09/21/2001
• Status: Active Grant

• Current Assignee: Garnet Biotherapeutics Incorporated
• Original Assignee: Garnet Biotherapeutics Incorporated
• Inventors: Ho, Tony W., Kopen, Gene C., Righter, William F., Rutkowski, J. Lynn, Wagner, Joseph
• Primary Examiner(s): Wityshyn, Michael G.
• Application Number:
  US11/054,824
• Publication Number:
  US 20050233452A1
• Time in Patent Office: 3,078 Days
• Field of Search: None
• US Class Current: 435/374
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2500/02 Atmosphere, e.g. low oxygen conditions
• CPC Class Codes: C12N 2500/32 Amino acids
• CPC Class Codes: C12N 2506/1353 from bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 5/0623 Stem cells
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 5/0669 Bone marrow stromal cells; Whole bone marrow isolated stem cells from bone marrow C12N5/0647, C12N5/0663

Abstract

Substantially homogenous cells populations which co-express CD49c, CD90 and telomerase are made. In one embodiment, humans suffering from a degenerative, traumatic, acute injury, cardiac or neurological condition are treated with the substantially homogenous cells populations which co-express CD49c, CD90 and telomerase. In another embodiment, committed progenitor cells are made are made by selecting from a cultured source of a cell population which co-express CD49c and CD90 and modifying the cell population. The committed progenitor cells can be employed to treat a human suffering from a degenerative, traumatic, acute injury, cardiac or neurological condition and formulate pharmaceutical compositions.

Claims

1. A method of producing an isolated cell population wherein greater than about 91% of the cells in said cell population co-express CD49c and CD90, comprising the steps of:

a) providing a source population of cells; and

,

b) culturing said source cell population at a seeding density of less than about 100 cells/cm² under a low oxygen condition.

2. The method of claim 1, wherein the source of the cell population is bone marrow.

3. The method of claim 2, wherein the bone marrow is human bone marrow.

4. The method of claim 1, wherein the low oxygen condition is less than about 15% oxygen.

5. The method of claim 4, wherein the low oxygen condition is less than about 10% oxygen.

6. The method of claim 1, wherein the low oxygen condition is about 5% oxygen.

7. The method of claim 2, further including lysing the red blood cells in the bone marrow prior to culturing the cells of the bone marrow.

8. The method of claim 2, further including fractionating the bone marrow prior to culturing the bone marrow.

9. The method of claim 8, wherein in the bone marrow is fractionated by passage through a density gradient.

10. The method of claim 8, wherein the bone marrow is fractionated by NH₂Cl lysis.

11. The method of claim 8, wherein the bone marrow is fractionated by fluorescent activated sorting.

12. The method of claim 8, wherein the bone marrow is fractionated by magnetic sorting.

13. The method of claim 1, wherein the cells which co-express CD49c and CD90 do not express CD34 and/or CD45.

14. The method of claim 1, wherein the cells isolated from the cultured source express at least one trophic factor selected from the group consisting of:

a) brain-derived neurotrophic factor (BDNF);

b) interleukin-6 (IL-6);

c) nerve growth factor (NGF);

d) macrophage chemoattractant protein-1 (MCP-1);

e) neurotrophin-3 (NT-3);

f) interleukin-7 (IL-7);

g) interleukin-11 (IL-11);

h) stem cell factor (SCF);

i) matrix metalloproteinase-9 (MMP-9); and

,

j) Cystatin-C.

22. The method of claim 1, wherein the low oxygen condition is between about 1% to about 10% oxygen.

23. The method of claim 1, wherein the low oxygen condition is between about 2% to about 7% oxygen.

15. A method of producing an isolated cell population wherein greater than about 91% of the cells in said cell population co-express CD49c and CD90, comprising the steps of:

a) providing a source population of cells; and

,

b) culturing said source cell population at a seeding density of less than about 100 cells/cm² under a low oxidative stress condition.

16. The method of claim 15, wherein the source of the cell population is bone marrow.

17. The method of claim 15, wherein the bone marrow is human bone marrow.

18. A method of producing an isolated cell population wherein greater than about 91% of the cells in said cell population co-express CD49c and CD90, comprising the steps of:

a) providing a source population of cells; and

,

b) culturing said source cell population at a seeding density of less than about 50 cells/cm² under a low oxidative stress condition.

19. A method of producing an isolated cell population wherein greater than about 91% of the cells in said cell population co-express CD49c and CD90, comprising the steps of:

a) providing a source population of cells; and

,

b) culturing said source cell population at a seeding density of less than about 30 cells/cm² under a low oxidative stress condition.

20. A method of producing an isolated cell population wherein greater than about 91% of the cells in said cell population co-express CD49c and CD90, comprising the steps of:

a) providing a source population of cells; and

,

b) culturing said source cell population at a seeding density of less than about 50 cells/cm² under a low oxygen condition.

24. The method of claim 20, wherein the source of the cell population is bone marrow.

25. The method of claim 24, wherein the bone marrow is human bone marrow.

26. The method of claim 20, wherein the low oxygen condition is less than about 15% oxygen.

27. The method of claim 26, wherein the low oxygen condition is less than about 10% oxygen.

28. The method of claim 20, wherein the low oxygen condition is about 5% oxygen.

29. The method of claim 20, wherein the low oxygen condition is between about 1% to about 10% oxygen.

30. The method of claim 20, wherein the low oxygen condition is between about 2% to about 7% oxygen.

31. The method of claim 24, further including lysing the red blood cells in the bone marrow prior to culturing the cells of the bone marrow.

32. The method of claim 24, further including fractionating the bone marrow prior to culturing the bone marrow.

33. The method of claim 32, wherein in the bone marrow is fractionated by passage through a density gradient.

34. The method of claim 32, wherein the bone marrow is fractionated by NH₂Cl lysis.

35. The method of claim 32, wherein the bone marrow is fractionated by fluorescent activated sorting.

36. The method of claim 32, wherein the bone marrow is fractionated by magnetic sorting.

37. The method of claim 20, wherein the cells which co-express CD49c and CD90 do not express CD34 and/or CD45.

38. The method of claim 20, wherein the cells isolated from the cultured source express at least one trophic factor selected from the group consisting of:

a) brain-derived neurotrophic factor (BDNF);

b) interleukin-6 (IL-6);

c) nerve growth factor (NGF);

d) macrophage chemoattractant protein-1 (MCP-1);

e) neurotrophin-3 (NT-3);

f) interleukin-7 (IL-7);

g) interleukin-11 (IL-11);

h) stem cell factor (SCF);

i) matrix metalloproteinase-9 (MMP-9); and

,

j) Cystatin-C.

21. A method of producing an isolated cell population wherein greater than about 91% of the cells in said cell population co-express CD49c and CD90, comprising the steps of:

a) providing a source population of cells; and

,

b) culturing said source cell population at a seeding density of less than about 30 cells/cm² under a low oxygen condition.

39. The method of claim 21, wherein the source of the cell population is bone marrow.

40. The method of claim 39, wherein the bone marrow is human bone marrow.

41. The method of claim 21, wherein the low oxygen condition is less than about 15% oxygen.

42. The method of claim 41, wherein the low oxygen condition is less than about 10% oxygen.

43. The method of claim 21, wherein the low oxygen condition is about 5% oxygen.

44. The method of claim 21, wherein the low oxygen condition is between about 1% to about 10% oxygen.

45. The method of claim 21, wherein the low oxygen condition is between about 2% to about 7% oxygen.

46. The method of claim 40, further including lysing the red blood cells in the bone marrow prior to culturing the cells of the bone marrow.

47. The method of claim 40, further including fractionating the bone marrow prior to culturing the bone marrow.

48. The method of claim 47, wherein the bone marrow is fractionated by passage through a density gradient.

49. The method of claim 47, wherein the bone marrow is fractionated by NH₂Cl lysis.

50. The method of claim 47, wherein the bone marrow is fractionated by fluorescent activated sorting.

51. The method of claim 47, wherein the bone marrow is fractionated by magnetic sorting.

52. The method of claim 21, wherein the cells which co-express CD49c and CD90 do not express CD34 and/or CD45.

53. The method of claim 21, wherein the cells isolated from the cultured source express at least one trophic factor selected from the group consisting of:

a) brain-derived neurotrophic factor (BDNF);

b) interleukin-6 (IL-6);

c) nerve growth factor (NGF);

d) macrophage chemoattractant protein-1 (MCP-1);

e) neurotrophin-3 (NT-3);

f) interleukin-7 (IL-7);

g) interleukin-11 (IL-11);

h) stem cell factor (SCF);

i) matrix metalloproteinase-9 (MMP-9); and

,

j) Cystatin-C.

54. A method of producing an isolated cell population wherein greater than about 91% of the cells in said cell population co-express CD49c and CD90 and maintain a constant population doubling rate for at least 8 population doublings, comprising the steps of:

a) providing a source population of cells; and

,

b) culturing said source cell population at a seeding density of less than about 100 cells/cm² under a low oxygen condition.

55. The method of claim 54, wherein the self-renewing colony forming units of cells maintain a constant population doubling rate for at least 16 population doublings.

56. The method of claim 55, wherein the self-renewing colony forming units of cells maintain a constant population doubling rate for at least 24 population doublings.

57. The method of claim 56, wherein the self-renewing colony forming units of cells maintain a constant population doubling rate for at least 32 population doublings.

58. The method of claim 57, wherein the self-renewing colony forming units of cells maintain a constant population doubling rate for at least 40 population doublings.

59. The method of claim 54, wherein the seeding density is less than about 50 cells/cm².

60. The method of claim 59, wherein the seeding density is less than about 30 cells/cm².

61. The method of claim 54, wherein the low oxygen condition is less than about 15% oxygen.

62. The method of claim 61, wherein the low oxygen condition is less than about 10% oxygen.

63. The method of claim 54, wherein the low oxygen condition is about 5% oxygen.

64. The method of claim 54, wherein the low oxygen condition is between about 1% to about 10% oxygen.

65. The method of claim 54, wherein the low oxygen condition is between about 2% to about 7% oxygen.

66. The method of claim 54, wherein the source of the cell population is bone marrow.

67. The method of claim 66, wherein the bone marrow is human bone marrow.

68. The method of claim 67, wherein the population doubling rate less than about 30 hours.