TARGET POPULATIONS OF OLIGODENDROCYTE PRECURSOR CELLS AND METHODS OF MAKING AND USING SAME

• US 20100158878A1
• Filed: 12/23/2009
• Published: 06/24/2010
• Est. Priority Date: 12/23/2008
• Status: Abandoned Application

• Current Assignee: BOCO Silicon Valley, Inc.
• Original Assignee: BOCO Silicon Valley, Inc.
• Inventors: Uchida, Nobuko, Capela, Alexandra
• Application Number:
  US12/646,228
• Publication Number:
  US 20100158878A1
• Time in Patent Office: Days
• US Class Current: 424/93.700
• 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 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/135 Platelet-derived growth factor [PDGF]
• CPC Class Codes: C12N 5/0622 Glial cells, e.g. astrocytes, oligodendrocytes; Schwann cells

Abstract

This application provides for enriched target populations oligodendrocyte precursor cells (OPCs) that can differentiate into oligodendrocytes. The target OPCs may be expanded and optionally subjected to conditions to induce their differentiation into oligodendrocytes. The target OPCs and their progeny are useful for the treatment of disease associated with demyelination of central nervous system axons.

Claims

1. An enriched population of oligodendrocyte precursor cells (OPCs) derived from neural or neural derived cells, wherein the population is enriched for target OPCs that are PDGFRα

⁺ and CD105⁻

, wherein at least 30% of the cells in the population are target OPCs.

2. The population of claim 1, where in the target cells are also PSA-NCAM^lo/−

.

3. The population of claim 1, where in the target cells are also A2B5^lo/−

.

4. The population of claim 1, where in the target cells are also CD133⁺.

5. The population of OPCs in claim 1, wherein at least 50% of the cells in the population are target OPCs.

6. The population of OPCs in claim 1, wherein at least 70% of the cells in the population are target OPCs.

7. The population of OPCs in claim 1, wherein at least 90% of the cells in the population are target OPCs.

8. The population of OPCs in claim 1, wherein the target OPCs are PDGFRα

⁺, CD105⁻

, CD133⁺, and A2B5⁻

.

9. The population of OPCs in claim 1, wherein the target OPCs are PDGFRα

⁺, CD105⁻

, CD133⁺, A2B5^lo/−

, and PSA-NCAM⁻

.

32. A method of treating a mammalian individual suffering from a disease associated with demyelination of central nervous system axons, comprising introducing the enriched population of OPCs of claim 1 to the mammalian individual in an amount effective to treat the disease.

33. The method of claim 32, wherein the mammalian individual is a human.

34. The method of claim 32, wherein the OPCs are administered to the mammalian individual by cell transplantation.

35. The method of claim 32, wherein the disease is multiple sclerosis, Pelizaeus-Merzbacher disease, cerebral palsy, radiation induced myelination disorders, acute disseminated encephalomyelitis, transverse myelitis, demyelinating genetic disease, spinal cord injury, virus-induced demyelination, Progressive Multifocal Leucoencephalopathy, Human Lymphotrophic T-cell Virus I (HTLVI)-associated myelopathy, or nutritional metabolic disorder.

36. The method of claim 32, wherein the disease is multiple sclerosis, Pelizaeus-Merzbacher disease, or cerebral palsy.

37. A method of treating a mammalian individual suffering from a disease associated with demyelination of central nervous system axons, comprising introducing the OPCs of claim 1 to the mammalian individual in an amount effective to treat the disease.

38. The method of claim 37, wherein the mammalian individual is a human.

39. The method of claim 37, wherein the OPCs are administered to the mammalian individual by cell transplantation.

40. The method of claim 37, wherein the disease is multiple sclerosis, acute disseminated encephalomyelitis, transverse myelitis, demyelinating genetic disease, spinal cord injury, virus-induced demyelination, Progressive Multifocal Leucoencephalopathy, Human Lymphotrophic T-cell Virus I (HTLVI)-associated myelopathy, or nutritional metabolic disorder.

45. A method of screening for compounds that affect a biological function of an enriched population of target oligodendrocyte precursor cells comprising:

(a) contacting an enriched population of target oligodendrocyte precursor cells obtained by the method of claim 1 with a test compound; and

(b) detecting a change in a biological function of the oligodendrocyte precursor cells.

46. The method of claim 45, wherein said target oligodendrocyte precursor cells are PDGFRα

⁺, CD105⁻

, A2B5^lo/−

, PSA-NCAM^lo/−

, and CD133⁺.

47. The method of claim 45, wherein said target oligodendrocyte precursor cells are PDGFRα

⁺, CD105⁻

, A2B5⁻

, and CD133⁺.

48. The method of claim 45, wherein said target oligodendrocyte precursor cells are PDGFRα

⁺, CD105⁻

, A2B5^lo/−

, PSA-NCAM⁻

, and CD133⁺.

49. The method of claim 45, wherein the change occurs in at least one of the characteristics selected from the group consisting of myelination, differentiation into oligodendrocytes, proliferation rate, cell migration, viability, gene expression, protein expression, protein levels in the culturing medium, dedifferentiation, growth characteristics, and cell morphology.

10. A method of producing population of cells enriched for target oligodendrocyte precursor cells (OPCs) from neural or neural derived cells, comprising contacting neural or neural derived cells with at least one reagent that binds to cell surface antigens expressed by the target OPCs, wherein the target OPCs are PDGFRα

⁺ and CD105⁻

, wherein the population of cells is enriched to contain at least 30% target OPC.

11. The method of claim 10, wherein the target OPCs are also PSA-NCAM^lo/−

.

12. The method of claim 10, where in the target cells are also A2B5^lo/−

.

13. The method of claim 10, where in the target cells are also CD133⁺.

14. The method of claim 10 further comprising negatively selecting for target OPCs by selectively removing nontarget cell populations.

15. The method of claim 10, wherein the target OPCs are PDGFRα

⁺, CD105⁻

, CD133⁺, and A2B5⁻

.

16. The method of claim 10, wherein the target OPCs are PDGFRα

⁺, CD105⁻

, CD133⁺, A2B5^lo/−

, and PSA-NCAM⁻

.

17. A method of increasing the number of target oligodendrocyte precursor cells (OPCs) in a cell culture comprising culturing a population of cells comprising at least 30% target OPCs to increase the number of target OPCs in the cell culture, wherein the target OPCs are PDGFRα

⁺ and CD105⁻

.

18. The method of claim 17, wherein the target OPCs are also PSA-NCAM^lo/−

.

19. The method of claim 17, where in the target cells are also A2B5^lo/−

.

20. The method of claim 17, where in the target cells are also CD133⁺.

21. The method of claim 17, wherein the target OPCs are PDGFRα

⁺, CD105⁻

, CD133⁺, and A2B5⁻

.

22. The method of claim 17, wherein the target OPCs are PDGFRα

⁺, CD105⁻

, CD133⁺, A2B5^lo/−

, and PSA-NCAM⁻

.

23. The method of claim 17 further comprising contacting the OPCs with an effective amount of at least one biological agent that is capable of increasing the number of OPCs.

24. The method of claim 23 wherein the biological agent is selected from the group consisting of leukemia inhibitory factor (LIF), epidermal growth factor (EGF), basic fibroblast growth factor (FGF-2;

bFGF), IGF-1, NT3, Shh, CTNF, PDGF-AA and combinations thereof.

41. A method of treating a mammalian individual suffering from a disease associated with demyelination of central nervous system axons, comprising introducing the differentiating or differentiated oligodendrocytes prepared according to the method of claim 21 to the mammalian individual in an amount effective to treat the disease.

42. The method of claim 41, wherein the mammalian individual is a human.

43. The method of claim 41, wherein the differentiating or differentiated oligodendrocytes are administered to the mammalian individual by cell transplantation.

44. The method of claim 41, wherein the disease is multiple sclerosis, acute disseminated encephalomyelitis, transverse myelitis, demyelinating genetic disease, spinal cord injury, virus-induced demyelination, Progressive Multifocal Leucoencephalopathy, Human Lymphotrophic T-cell Virus I (HTLVI)-associated myelopathy, or nutritional metabolic disorder.

25. A method for proliferating oligodendrocyte precursor cells (OPCs) comprising:

proliferating OPCs in serum-free culture medium containing one or more predetermined growth factors effective for inducing OPC proliferation, wherein;

(a) the population comprises OPCs which are PDGFRα

⁺ and CD105⁻

; and

(b) in the presence of differentiation-inducing conditions, the cells produce progeny cells that differentiate into oligodendrocytes.

26. The method of claim 25, wherein the OPCs are also PSA-NCAM^lo/−

.

27. The method of claim 25, where in the target cells are also A2B5^lo/−

.

28. The method of claim 25, where in the target cells are also CD133⁺.

29. The method of claim 25 further comprising subjecting the OPCs to culture conditions that induce oligodendrocyte differentiation to produce differentiating and differentiated oligodendrocytes.

30. A composition comprising the oligodendrocyte precursor cells (OPCs) produced by the method of claim 25.

31. A composition comprising the differentiating or differentiated oligodendrocytes produced by the method of claim 25.

50. A method of producing a population enriched for oligodendrocyte precursor cells comprising:

(a) contacting neural or neural derived cells comprising one or more multipotent central nervous system stem cell with an antibody that specifically binds to PDGFRα

; and

(b) selecting said neural or neural derived cells that are PDGFRα

^hi, wherein the selected cells are enriched for oligodendrocyte precursor cells as compared with the neural or neural derived cells.

51. The method of claim 50, wherein said neural or neural derived cells are obtained from a neuro sphere culture or an adherent culture.

52. The method of claim 50, wherein the method further comprises the step of eliminating those cells that are PDGFRα

^lo/med.

53. The method of claim 50, wherein the method further comprises the step of eliminating cells that are CD105⁻

.