Differentiation of whole bone marrow

• US 20030082160A1
• Filed: 09/24/2002
• Published: 05/01/2003
• Est. Priority Date: 10/25/2001
• Status: Active Grant

• Current Assignee: Cedars-Sinai Medical Center (Cedars Sinai Health System)
• Original Assignee: Cedars-Sinai Medical Center (Cedars Sinai Health System)
• Inventors: Yu, John S., Ehtesham, Moneeb, Kabos, Peter
• Granted Patent: US 7,129,034 B2
• Time in Patent Office: Days
• US Class Current: 424/93.21
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 9/00 Drugs for disorders of the cardiovascular system
• 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 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2506/11 from blood or immune system cells
• CPC Class Codes: C12N 2506/1353 from bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

A method is described for generating a clinically significant volume of neural progenitor cells from whole bone marrow. A mass of bone marrow cells may be grown in a culture supplemented with fibroblast growth factor-2 (FGF-2) and epidermal growth factor (EGF). Further methods of the present invention are directed to utilizing the neural progenitor cells cultured in this fashion in the treatment of various neuropathological conditions, and in targeting delivery of cells transfected with a particular gene to diseased or damaged tissue.

Claims

1. A method to generate a clinically significant volume of neural progenitor cells, the method comprising:

providing a mass of cells obtained from a mammal including at least one stem cell; and

culturing the at least one stem cell in a medium comprising fibroblast growth factor-2 (FGF-2) and epidermal growth factor (EGF) to produce a clinically significant volume of neural progenitor cells.

2. The method of claim 1, wherein the mass of cells is obtained from bone marrow of the mammal.

3. The method of claim 1, wherein the mass of cells is obtained from an adult mammal.

4. The method of claim 1, wherein the mass of cells is obtained from a fetus.

5. The method of claim 1, wherein the mass of cells is obtained from a brain of the mammal.

6. The method of claim 1, wherein the medium is Dulbecco'"'"'s modified Eagle medium (DMEM).

7. The method of claim 1, wherein the medium further comprises a supplement.

8. The method of claim 7, wherein the supplement is selected from the group consisting of B27, N2, and combinations thereof.

9. The method of claim 1, wherein the medium further comprises an additional compound.

10. The method of claim 9, wherein the additional compound is selected from the group consisting of interleukin-3 (IL-3), stem cell factor-1 (SCF-1), sonic hedgehog (Shh), fms-like tyrosine kinase-3 (Flt3) ligand, leukemia inhibitory factor (LIF), and combinations thereof.

11. The method of claim 1, wherein the medium further comprises an antibiotic.

12. The method of claim 11, wherein the antibiotic is selected from the group consisting of penicillin, streptomycin, and combinations thereof.

13. The method of claim 1, wherein clusters of the neural progenitor cells develop in the medium, and the method further comprises separating at least one cluster from the medium.

14. The method of claim 13, further including subculturing the at least one cluster.

15. The method of claim 14, wherein subculturing the at least one cluster further comprises culturing the at least one cluster in a medium comprising FGF-2 and EGF.

16. A method to treat a neuropathologic condition, the method comprising:

providing a mass of cells obtained from a mammal including at least one stem cell;

culturing the at least one stem cell in a medium comprising fibroblast growth factor-2 (FGF-2) and epidermal growth factor (EGF) to produce a neural progenitor cell; and

administering the neural progenitor cell to a mammal.

17. The method of claim 16, wherein the mass of cells is obtained from bone marrow of the mammal.

18. The method of claim 16, wherein the mass of cells is obtained from an adult mammal.

19. The method of claim 16, wherein the mass of cells is obtained from a fetus.

20. The method of claim 16, wherein the mass of cells is obtained from a brain of the mammal.

21. The method of claim 16, wherein the medium is Dulbecco'"'"'s modified Eagle medium (DMEM).

22. The method of claim 16, wherein the medium further comprises a supplement.

23. The method of claim 22, wherein the supplement is selected from the group consisting of B27, N2, and combinations thereof.

24. The method of claim 16, wherein the medium further comprises an additional compound.

25. The method of claim 24, wherein the additional compound is selected from the group consisting of interleukin-3 (IL-3), stem cell factor-1 (SCF-1), sonic hedgehog (Shh), fms-like tyrosine kinase-3 (Flt3) ligand, leukemia inhibitory factor (LIF), and combinations thereof.

26. The method of claim 16, wherein the medium further comprises an antibiotic.

27. The method of claim 26, wherein the antibiotic is selected from the group consisting of penicillin, streptomycin, and combinations thereof.

28. The method of claim 16, wherein clusters of neural progenitor cells develop in the medium, and the method further comprises separating at least one cluster from the medium.

29. The method of claim 28, further including subculturing the at least one cluster.

30. The method of claim 29, wherein subculturing the at least one cluster further comprises culturing the at least one cluster in a medium comprising FGF-2 and EGF.

31. The method of claim 16, wherein the neuropathologic condition is selected from the group consisting of brain tumor, brain ischemia, stroke, spinal cord injury, and neurodegenerative disorders.

32. The method of claim 16, wherein administering the neural progenitor cell further comprises administering a clinically significant volume of neural progenitor cells.

33. The method of claim 32, wherein the clinically significant volume of neural progenitor cells is sufficient to treat the neuropathologic condition.

34. A method to deliver a biological compound, the method comprising:

providing a mass of cells obtained from a mammal including at least one stem cell;

culturing the at least one stem cell in a medium comprising fibroblast growth factor-2 (FGF-2) and epidermal growth factor (EGF) to produce a neural progenitor cell;

causing the neural progenitor cell to carry a gene that effects local production of the biological compound; and

administering the neural progenitor cell to a mammal.

35. The method of claim 34, wherein the mass of cells is obtained from bone marrow of the mammal.

36. The method of claim 34, wherein the mass of cells is obtained from an adult mammal.

37. The method of claim 34, wherein the mass of cells is obtained from a fetus.

38. The method of claim 34, wherein the mass of cells is obtained from a brain of the mammal.

39. The method of claim 34, wherein the medium is Dulbecco'"'"'s modified Eagle medium (DMEM).

40. The method of claim 34, wherein the medium further comprises a supplement.

41. The method of claim 40, wherein the supplement is selected from the group consisting of B27, N2, and combinations thereof.

42. The method of claim 34, wherein the medium further comprises an additional compound.

43. The method of claim 42, wherein the additional compound is selected from the group consisting of interleukin-3 (IL-3), stem cell factor-1 (SCF-1), sonic hedgehog (Shh), fms-like tyrosine kinase-3 (Flt3) ligand, leukemia inhibitory factor (LIF), and combinations thereof.

44. The method of claim 34, wherein the medium further comprises an antibiotic.

45. The method of claim 44, wherein the antibiotic is selected from the group consisting of penicillin, streptomycin, and combinations thereof.

46. The method of claim 34, wherein clusters of neural progenitor cells develop in the medium, and the method further comprises separating at least one cluster from the medium.

47. The method of claim 46, further including subculturing the at least one cluster.

48. The method of claim 47, wherein subculturing the at least one cluster further comprises culturing the at least one cluster in a medium comprising FGF-2 and EGF.

49. The method of claim 34, wherein the biological compound is selected from the group consisting of cytokines, brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4), neurotrophin-5 (NT-5), basic fibroblast growth factor (bFGF), insulin-like growth factor-1 (IGF-1), glial-derived neurotrophic factor (GDNF), amyloid precursor proteins, protease nexin-1, and combinations thereof.

50. The method of claim 34, wherein the neural progenitor cell is administered to treat a neuropathologic condition.

51. The method of claim 50, wherein the neuropathological condition is selected from the group consisting of brain tumor, brain ischemia, stroke, spinal cord injury, and neurodegenerative disorders.

52. The method of claim 34, wherein administering the neural progenitor cell further comprises administering a clinically significant volume of neural progenitor cells.