Novel mammalian multipotent neural stem cells and compositions, methods of preparation and methods of administration thereof

• US 20030148513A1
• Filed: 01/14/2003
• Published: 08/07/2003
• Est. Priority Date: 01/14/2002
• Status: Active Grant

• Current Assignee: Board of Trustees of The University of Illinois
• Original Assignee: Board of Trustees of The University of Illinois
• Inventors: Sugaya, Kiminobu, Qu, Tingyu, Pulido, Jose S.
• Granted Patent: US 7,618,621 B2
• Time in Patent Office: Days
• US Class Current: 435/368
• CPC Class Codes: A61K 31/505 Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
• CPC Class Codes: A61K 31/519 ortho- or peri-condensed with heterocyclic rings
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• 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 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• 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 25/08 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 25/18 Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• 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 27/00 Drugs for disorders of the senses
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: A61P 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/00 : Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• 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 2500/90 : Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2500/99 : Serum-free medium
• CPC Class Codes: C12N 2501/06 : Anti-neoplasic drugs, anti-retroviral drugs, e.g. azacytidine, cyclophosphamide
• 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 2501/15 : Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/165 : Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2501/91 : Heparin
• CPC Class Codes: C12N 2501/998 : Proteins not provided for elsewhere
• CPC Class Codes: C12N 2501/999 : Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2502/08 : cells of the nervous system
• CPC Class Codes: C12N 2506/1353 : from bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 5/0621 : Eye cells, e.g. cornea, iris pigmented cells photoreceptors C12N5/062
• CPC Class Codes: C12N 5/0623 : Stem cells
• CPC Class Codes: C12N 5/0662 : Stem cells

Abstract

This invention relates to novel mammalian multipotent neural stem cells (MNSCs), compositions thereof, and methods of preparing and administering the cells to diseased, aged or damaged tissue such that the cells properly migrate and differentiate and a neurological or corporal deficit is improved or remedied as a result.

Claims

43. A method of making a retinal cell, the method comprising contacting a neural stem cell before or during differentiation with an effective amount of growth factor selected from the group consisting of TGF-b3, IGF-1 and CNTF, for an effective period, wherein the growth factor-contacted neural stem cells become capable of differentiating into retinal cells.

1. A method of preparing one or a plurality of mammalian multipotent stem cells that can forms a cluster comprising the steps of

a. isolating mammalian neural stem cells derived from mammalian neural tissue containing at least one neural stem cell,

b. culturing said cells in a cell culture media under conditions that permit proliferation of the neural stem cells in vitro,

c. harvesting the one or plurality of mammalian multipotent stem cells that can form a cluster produced in (b),

wherein said one or a plurality of mammalian multipotent stem cells that can form a cluster is adapted for proliferation, migration and differentiation in mammalian tissue when introduced thereto.

2. The method according to claim 1 further comprising the step of culturing said cells in a cell culture media comprising an-effective amount of substituted deoxynucleotide or deosynucleoside compound for an effective period and at least one growth factor and wherein cells are cultured in an uncoated flask or a flask that has been treated to repel the cells.

3. The method of claim 2 wherein the substituted deoxynucleotide or deoxynucleoside compound is a halo-substituted substituted deoxynucleotide or halo-substituted deoxynucleoside.

4. The method of claim 2 wherein the substituted deoxynucleotide or deoxynucleoside compound is a bromodeoxynucleotide or bromodeoxynucleoside.

5. The method of claim 2 wherein the substituted deoxynucleotide or deoxynucleoside compound is an iododeoxynucleotide or iododeoxynucleoside.

6. The method of claim 2 wherein the substituted deoxynucleotide or deoxynucleoside compound is an alkyl-substituted deoxynucleotide or alkyl substituted deoxynucleoside.

7. The method of claim 2 wherein the substituted deoxynucleotide or deoxynucleoside compounds is a methyldeoxynucleotide or methyldeoxynucleoside.

8. The method of claim 1 or 2 wherein the growth factor in the culture media prepared (b) is fibroblast growth factor, epidermal growth factor or combinations thereof.

9. The method of claim 1 or 2 wherein said growth factor in the culture media prepared in (b) is fibroblast growth factor or epidermal growth factor.

10. The method of claim 1 or 2 wherein said growth factor in th culture media prepared in (b) is fibroblast growth factor.

11. The method of claim 1 or 2 wherein the growth factor in the culture media prepared in (b) is epidermal growth factor.

12. The method of claim 1 or 2 wherein the culture medium of step (b) further comprises heparin.

13. The method of claim 1 or 2 wherein said mammalian neural tissue is obtained from a zygote, blastocyst, embryo, fetus, infant juvenile or adult.

14. The method of claim 1 or 2 wherein said mammalian neural tissue is obtained from a human.

15. One or a plurality of mammalian multipotent stem cells that can form a cluster produced according to the method of claim 1 or 2.

16. The plurality of mammalian multipotent stem cells that can form a cluster of claim 15 comprising less than about 50 percent differentiated neural stem cells.

17. The plurality of mammalian multipotent stem cells that can form a cluster of claim 15 comprising less than about 25 percent differentiated neural stem cells.

18. The plurality of mammalian multipotent stem cells that can form a cluster of claim 15 comprising less than about 10 percent differentiated neural stem cells.

19. The plurality of mammalian multipotent stem cells that can form a cluster of claim 15 comprising less than about 5 percent differentiated neural stem cells.

20. The plurality of mammalian multipotent stem cells that can form a cluster of claim 15 comprising less than about 1 percent differentiated neural stem cells.

21. A method for treating a patient having a neurological deficit or corporal deficit with one or a plurality of mammalian multipotent stem cells that can form a 15 cluster produced according to the method of claim 1 or 2 comprising the step of administering said mammalian multipotent stem cells to an animal having a neurological deficit or corporal deficit.

22. A composition for cell or tissue regeneration comprising, as an active ingredient, the one or a plurality of mammalian multipotent stem cells that can form a cluster produced according to the method of claim 1 or 2.

23. A method of regenerating cells or tissue comprising the step of administering the preparation of claim 22 to an animal in need thereof.

24. A pharmaceutical composition for treating a neurological deficit or corporal deficit comprising, as an active ingredient, one or a plurality of mammalian multipotent stem cells that can form a cluster produced according to the method of claim 1 or 2.

25. The pharmaceutical composition according to claim 24 further comprising a pharmaceutically acceptable carrier.

26. A method for treating a patient having a neurological deficit or corporal deficit, the method comprising:

a. administering an effective amount of the one or plurality of mammalian multipotent stem cells that can form a cluster of claim 24 or 25 to the patient with a neurological deficit or corporal deficit;

wherein the one or a plurality of mammalian multipotent stem cells that can form a cluster migrates to an area of tissue damage, differentiates in a tissue-specific manner and function in a manner that reduces the neurological or corporal deficit.

27. A method of administering one or a plurality of mammalian multipotent stem cells that can form a cluster produced by the method of claim 1 or 2 that is by injecting with a syringe, inserting with a catheter or surgical implantation.

28. The method of claim 23 or 26 wherein the one or plurality of mammalian multipotent stem cells that can form a cluster is injected with a syringe into a body, cavity that is fluidly-connected to the area of neurological or corporal deficit.

29. The method of claim 23 or 26 wherein the composition of one or a plurality of mammalian multipotent stem cells that can form a cluster is inserted with a catheter into a body cavity that is fluidly-connected to the area of neurological or corporal deficit.

30. The method of claim 23 or 26 wherein the composition of one or a plurality of mammalian multipotent stem cells that can form a cluster is surgically implanted into a body cavity that is fluidly-connected to the area of neurological or corporal deficit.

31. The method of claim 23 or 26 wherein the composition of one or a plurality;

of mammalian multipotent stem cells that can form a cluster is injected with a syringe to the area of neurological or corporal deficit.

32. The method of claim 23 or 26 wherein the composition of one or a plurality of mammalian multipotent stem cells that can form a cluster is inserted with a catheter to the area of neurological or corporal deficit.

33. The method of claim 23 or 26 wherein the composition of one or a plurality of mammalian multipotent stem cells that can form a cluster is surgically implanted to the area of neurological or corporal deficit.

34. The method of claim 23 or 26 wherein the composition of one or a plurality of mammalian multipotent stem cells that can form a cluster is introduced into a brain ventricle.

35. The method of claim 23 or 26 wherein the composition of one or a plurality of mammalian multipotent stem cells that can form a cluster is introduced into the brain ventricle by injection using a syringe.

36. The method of claim 23 or 26 wherein the composition of one or a plurality of mammalian multipotent stem cells that can form a cluster is introduced into the brain ventricle by insertion using a catheter.

37. The method of claim 23 or 26 wherein the composition of one or a plurality of mammalian multipotent stem cells that can form a cluster is introduced into the brain ventricle by surgical implantation.

38. The method of claim 23 or 26 wherein the composition of a plurality of mammalian multipotent stem cells that can form a cluster remains substantially intact during administration.

39. The method of claim 26, wherein the neurological deficit is caused by a neurodegenerative disease, a traumatic injury, a neurotoxic injury, ischemia, a developmental disorder, a disorder affecting vision, an injury or disease of the spinal cord, a demyelinating disease, an autoimmune disease, an infection, or an inflammatory disease.

40. The method of claim 26, wherein the corporal deficit is caused by corporal disease, disorder, injury, trauma, malfunction, degeneration or loss.

41. The method of claim 23 or 26, wherein the composition of one or a plurality of mammalian multipotent stem cells that can form a cluster is introduced systemically.

42. A method of making a terminally differentiated cell, the method comprising:

(a) administering the one or plurality of mammalian multipotent stem cells that can form a cluster prepared according to claim 1 or 2 to a patient with a neurological deficit or corporal deficit;

wherein the one or plurality of mammalian multipotent stem cells that can form a cluster migrates to an area of tissue damage, differentiates in a tissue-specific manner and functions in a manner that reduces the neurological deficit or corporal deficit.

44. The method of claim 43 wherein the neural stem cell is also contacted with a combination of two or more growth factors selected from the group consisting of TGF-b3, IGF-1 and CNTF for an effective period, wherein the growth factor-contacted neural stem cells become capable of differentiating into retinal cells.

45. A method for treating a patent having a neurological deficit, the method comprising the step of:

(a) administering the retinal cell of claim 43 or 44 to the patient;

wherein the retinal cells are capable of migrating to an area of tissue damage, differentiating in a tissue-specific manner and functioning in a manner that reduces the neurological deficit.

46. Terminally differentiated cells produced from the one or a plurality of mammalian multipotent stem cells that can form a cluster of claim 1 or 2.

47. A method for treating a patient having a neurological deficit or corporal deficit with terminally differentiated cells of claim 46, comprising the step of administering said terminally differentiated cells to an animal having a neurological deficit or corporal deficit.

45-1. Cells produced according to the methods of claim 43 or 44.