Method of treating a nervous system injury with cultured schwann cells

• US 6,033,660 A
• Filed: 05/10/1995
• Issued: 03/07/2000
• Est. Priority Date: 05/10/1995
• Status: Expired due to Term

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Chen, Jian, Mather, Jennie P., Li, Ronghao
• Primary Examiner(s): Witz, Jean C.
• Application Number:
  US08/438,862
• Time in Patent Office: 1,763 Days
• Field of Search: 435/240.1, 435/240.2, 435/240.3, 424/93.7
• US Class Current: 424/93.7
• 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: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/195 Heregulin, neu differentiation factor
• CPC Class Codes: C12N 2501/40 Regulators of development
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 5/0622 Glial cells, e.g. astrocytes, oligodendrocytes; Schwann cells

Abstract

A method for enhancing the survival and/or proliferation of Schwann cells (especially human Schwann cells) in cell culture is disclosed which involves culturing the cells in serum free culture medium comprising gas6 and other mitogenic agents, such as heregulin and forskolin. The culturing step is generally preceded by a pre-incubation period wherein nerve tissue comprising the Schwann cells is cultured under appropriate conditions and for a period of time such that demyelination occurs. The isolated Schwann cells can be used as cellular prostheses to treat patients with nervous system injuries. The invention also provides a cell culture medium for culturing Schwann cells.

Claims

1. A method of treating a nervous system injury in a mammal comprising administering to the mammal human Schwann cells which have been cultured in serum free culture medium, wherein the culture medium comprises a nutrient solution for growing mammalian cells supplemented with a first mitogenic agent which is a Rse/Axl receptor activator and a second mitogenic agent, wherein the first and second mitogenic agents are each present in the culture medium in an amount effective to enhance survival or proliferation of the Schwann cells, wherein said human Schwann cells are administered in an amount effective to promote nervous system repair at the site of said injury.

3. The method according to claim 1, wherein the second mitogenic agent is an erbB receptor activator.

4. The method according to claim 1, wherein the second mitogenic agent is an agent which elevates cAMP levels.

5. The method according to claim 4, wherein the agent which elevates cAMP levels is forskolin.

6. The method according to claim 1, wherein said culture medium additionally comprises a molecule or composition which provides Fe ions to the Schwann cells, wherein the molecule or composition is present in the culture medium in an amount effective to enhance survival or proliferation of the Schwann cells cultured therein.

7. The method according to claim 6, wherein said molecule or composition which provides Fe ions to the Schwann cells is transferrin.

8. The method according to claim 1, wherein the second mitogenic agent is insulin or an insulin like growth factor, and the insulin or insulin like growth factor is present in the culture medium in an amount effective to enhance survival or proliferation of Schwann cells cultured therein.

9. The method according to claim 1, wherein said culture medium additionally comprises Vitamin E, wherein the concentration of Vitamin E in the culture medium is in the range from 0.1 μ

g/ml to 100 μ

g/ml.

10. The method according to claim 1, wherein said culture medium additionally comprises a protease inhibitor, wherein the concentration of protease inhibitor in the culture medium is in the range from 1 μ

g/ml to 500 μ

g/ml.

2. A method of treating a nervous system injury in a mammal comprising administering to the mammal a composition comprising human Schwann cells which have been cultured in serum free culture medium, wherein the culture medium comprises a nutrient solution for growing mammalian cells supplemented with a first mitogenic agent which is a Rse/Axl receptor activator and a second mitogenic agent, wherein the first and second mitogenic agents are each present in the culture medium in an amount effective to enhance survival or proliferation of the Schwann cells, and a pharmaceutically acceptable carrier, wherein said composition is essentially free of serum and is administered in an amount effective to promote nervous system repair at the site of said injury.

11. The method according to claim 2, wherein said composition further comprises a mitogenic agent.

12. The method according to claim 11, wherein said mitogenic agent is a Rse receptor activator.

13. The method according to claim 12, wherein said Rse receptor activator is gas6.

14. The method according to claim 2, wherein said composition comprises about 5% serum.

15. The method according to claim 2, wherein said composition comprises about 1% serum.

16. The method according to claim 2, wherein said composition comprises about 0.1% serum.

17. The method according to claim 2, wherein said composition is free of serum.

18. A method of treating a nervous system injury in a mammal comprising:

a) culturing human Schwann cells in a serum free culture medium comprising a nutrient solution for growing mammalian cells supplemented with a first mitogenic agent which is a Rse/Axl receptor activator and a second mitogenic agent, wherein the first and second mitogenic agents are each present in the culture medium in an amount effective to enhance survival or proliferation of the Schwann cells; and

b) administering said cultured Schwann cells to a mammal in an amount effective to promote nervous system repair at the site of said injury.

19. A method of treating a nervous system injury in a mammal comprising:

administering to the mammal a serum free composition comprising human Schwann cells, wherein said Schwann cells comprise a first mitogen receptor which is a Rse/Axl receptor and a second mitogen receptor, wherein the first and second receptors each have been activated in an amount and under conditions effective to enhance survival or proliferation of the Schwann cells, wherein said composition is administered in an amount effective to promote nervous system repair at the site of said injury.

20. A method of treating a nervous system injury in a mammal comprising:

administering to the mammal a composition comprising human Schwann cells and a serum free medium comprising a nutrient solution for growing mammalian cells supplemented with a first mitogenic agent which is a Rse/Axl receptor activator and a second mitogenic agent, wherein the first and second mitogenic agents are each present in the medium in an amount effective to enhance survival or proliferation of human Schwann cells, wherein said composition is administered in an amount effective to promote nervous system repair at the site of said injury.

21. A method of treating a nervous system injury in a mammal comprising:

administering to the mammal a serum free composition comprising human Schwann cells comprising an first activated mitogen receptor which is a Rse/Axl receptor and a second activated mitogen receptor, wherein said first and second receptors have been activated in an amount effective to enhance survival or proliferation of the Schwann cells, wherein said composition is administered in an amount effective to promote nervous system repair at the site of said injury.