Expansion of neural stem cells with LIF

• US 20050214941A1
• Filed: 03/16/2005
• Published: 09/29/2005
• Est. Priority Date: 03/16/2004
• Status: Abandoned Application

• Current Assignee: Theradigm, Inc.
• Original Assignee: Theradigm, Inc.
• Inventors: Bhonsale, Smita, Vanguri, Padmavathy
• Application Number:
  US11/083,231
• Publication Number:
  US 20050214941A1
• Time in Patent Office: Days
• US Class Current: 435/368
• 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: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/08 Antiepileptics; Anticonvulsants
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• 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: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2533/32 Polylysine, polyornithine
• CPC Class Codes: C12N 2533/52 Fibronectin; Laminin
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

The present invention encompasses methods and compositions for enhancing the growth of neural stem cells (NSCs).

Claims

1. A composition comprising an in vitro adherent culture comprising a Neural Stem Cell (NSC), wherein said NSC proliferates in the presence of LIF while maintaining multipotentiality of said NSC.

2. The composition of claim 1, wherein said NSC adheres to a surface coated with polyornithine and fibronectin.

3. The composition of claim 1, wherein said NSC is derived from a human.

4. The composition of claim 1, wherein exogenous genetic material has been introduced into said NSC.

5. A method for the in vitro expansion and maintenance of the multipotentiality of a Neural Stem Cell (NSC), said method comprising culturing said NSC as an adherent population on a coated surface in the presence of LIF.

6. The method of claim 5, wherein said NSC adheres to a surface coated with polyornithine and fibronectin.

7. The method of claim 5, wherein said NSC is derived from a human.

8. The method of claim 5, wherein exogenous genetic material has been introduced into said NSC.

9. A method for the in vitro expansion and maintenance of the multipotentiality of an NSC, said method comprising culturing said NSC as an adherent population on a coated surface in the presence of LIF, wherein the expression of MHC class II molecule in said NSC is regulated by said method.

10. A method for the in vitro expansion and maintenance of the multipotentiality of a Neural Stem Cell (NSC), wherein the expression of MHC class II molecule is reduced in said NSC when compared to an otherwise identical NSC cultured in the continuous presence of LIF, said method comprising:

a) culturing said NSC as an adherent population on a coated surface in the presence of LIF for a period of time, then

b) removing LIF from the culture, and

c) culturing said NSC as an adherent population on a coated surface in the absence of LIF for a period of time.

11. The method of claim 10, wherein said NSC is cultured in the presence of LIF for about 7 days.

12. The method of claim 10, wherein said NSC is cultured in the absence of LIF for about 7 days.

13. The method of claim 10, wherein following (c), said NSC exhibits a doubling rate of about 28-36 hours.

14. An isolated Neural Stem Cell (NSC) prepared by a method of culturing said NSC comprising:

a) culturing said NSC as an adherent population on a coated surface in the presence of LIF for a period of time, then

b) removing LIF from the culture, and

c) culturing said NSC as an adherent population on a coated surface in the absence of LIF for a period of time.

15. The isolated NSC of claim 14, wherein said NSC exhibits a doubling rate of about 28-36 hours.

16. The isolated NSC of claim 14, wherein said NSC exhibits a reduced level of MHC class II molecule expression compared to the level of MHC class II molecule expression on an otherwise identical NSC cultured in the continuous presence of LIF.

17. The isolated NSC of claim 14, wherein said NSC is derived from a human.

18. The isolated NSC of claim 14, wherein exogenous genetic material has been introduced into said NSC.

19. A method of treating a human patient having a disease, disorder or condition of the central nervous system, the method comprising:

i) obtaining an isolated Neural Stem Cell (NSC),

ii) culturing said NSC as an adherent population on a coated surface in the presence of LIF for a period of time,

iii) removing LIF from the culture,

iv) culturing said NSC as an adherent population on a coated surface in the absence of LIF for a period of time, and

v) administering said cultured NSC to the central nervous system of said human patient.

20. The method of claim 19, wherein said disease, disorder or condition of the central nervous system is selected from the group consisting of a genetic disease, brain trauma, Huntington'"'"'s disease, Alzheimer'"'"'s disease, Parkinson'"'"'s disease, spinal cord injury, stroke, multiple sclerosis, cancer, CNS lysosomal storage diseases and head trauma, epilepsy.

21. The method of claim 19, wherein said disease, disorder or condition is injury to the tissue or cells of said central nervous system.

22. The method of claim 19, wherein said disease, disorder or condition is a brain tumor.

23. The method of claim 19, wherein said cultured NSC administered to said central nervous system remains present and/or replicates in said central nervous system.

24. The method of claim 19, wherein prior to administering said NSC, said NSC is further cultured in vitro in a differentiation medium.

25. The method of claim 19, wherein prior to administering said NSC, said NSC is genetically modified.

26. A composition comprising an isolated Neural Stem Cell (NSC) and a biologically compatible lattice, wherein said NSC is prepared by a method comprising:

a) culturing said NSC as an adherent population on a biologically compatible lattice in the presence of LIF for a period of time, then

b) removing LIF from the culture, and

c) culturing said NSC as an adherent population on a biologically compatible lattice in the absence of LIF for a period of time.

27. The composition of claim 26, wherein the lattice comprises polymeric material.

28. The composition of claim 26, wherein the polymeric material is formed of polymer fibers as a mesh or sponge.

29. The composition of claim 26, wherein the polymeric material comprises monomers selected from the group of monomers consisting of glycolic acid, lactic acid, propyl fumarate, caprolactone, hyaluronan, hyaluronic acid and combinations thereof.

30. The composition of claim 26, wherein the polymeric material comprises proteins, polysaccharides, polyhydroxy acids, polyorthoesters, polyanhydrides, polyphosphazenes, synthetic polymers or combinations thereof.

31. The composition of claim 26, wherein the polymeric material is a hydrogel formed by crosslinking of a polymer suspension having the cells dispersed therein.

32. The composition of claim 26, wherein said biologically compatible lattice is further coated with polyornithine.

33. The composition of claim 26, wherein said biologically compatible lattice is further coated with fibronectin.

34. The composition of claim 26, wherein said biologically compatible lattice is further coated with polyornithine and fibronectin.

35. A method for the in vitro expansion and maintenance of the multipotentiality of a Neural Stem Cell (NSC), said method comprising:

a) culturing said NSC as an adherent population on a biologically compatible lattice in the presence of LIF for a period of time, then

b) removing LIF from the culture, and

c) culturing said NSC as an adherent population on a biologically compatible lattice in the absence of LIF for a period of time.

36. The method of claim 35, wherein said biologically compatible lattice is further coated with polyornithine.

37. The method of claim 35, wherein said biologically compatible lattice is further coated with fibronectin.

38. The method of claim 35, wherein said biologically compatible lattice is further coated with polyornithine and fibronectin.