Differentiation of bone marrow cells into neuronal cells and uses therefor

• US 20030203484A1
• Filed: 08/06/2002
• Published: 10/30/2003
• Est. Priority Date: 02/11/2000
• Status: Active Grant

• Current Assignee: Philadelphia Health and Education Corporation, Rutgers University
• Original Assignee: Philadelphia Health and Education Corporation, University of Medicine and Dentistry of New Jersey (State of New Jersey)
• Inventors: Prockop, Darwin J., Black, Ira B., Woodbury, Dale L., Schwarz, Emily
• Granted Patent: US 7,279,331 B2
• Time in Patent Office: Days
• US Class Current: 435/368
• CPC Class Codes: A61K 31/045 Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
• CPC Class Codes: A61K 31/10 Sulfides; Sulfoxides; Sulfones
• CPC Class Codes: A61K 31/19 Carboxylic acids, e.g. valproic acid salicylic acid A61K31/60
• CPC Class Codes: A61K 31/198 Alpha-amino acids, e.g. alanine or edetic acid [EDTA] betaine A61K31/205; proline A61K31/401; tryptophan A61K31/405; histidine A61K31/4172; peptides not degraded to individual amino acids A61K38/00
• CPC Class Codes: A61K 31/375 Ascorbic acid, i.e. vitamin C; Salts thereof
• 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 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 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 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/08 Antiepileptics; Anticonvulsants
• 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: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/815 Dopamine
• CPC Class Codes: C12N 2501/999 : Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2506/1353 : from bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 5/0619 : Neurons

Abstract

The present invention relates to methods of inducing differentiation of mammalian bone marrow stromal cells into neuronal cells by contacting marrow stromal cells with a neuronal differentiation-inducing compounds. Neuronal differentiation-inducing compounds of the invention include anti-oxidants such as, but not limited to, beta-mercaptoethanol, dimethylsulfoxide, butylated hydroxyanisole, butylated hydroxytoluene, ascorbic acid, dimethylfumarate, and n-acetylcysteine. Once induced to differentiate into neuronal cells, the cells can be used for cell therapy, gene therapy, or both, for treatment of diseases, disorders, or conditions of the central nervous system.

Claims

1. A method of inducing differentiation of an isolated marrow stromal cell into a neuronal cell, said method comprising contacting said isolated marrow stromal cell with at least one neuronal differentiation-inducing compound, thereby inducing differentiation of said isolated marrow stromal cell into a neuronal cell.

2. The method of claim 1, wherein said isolated marrow stromal cell is a human cell.

3. The method of claim 1, wherein said neuronal differentiation-inducing compound is a trophic factor.

4. The method of claim 1, wherein said neuronal differentiation-inducing compound is a growth factor.

5. The method of claim 4, wherein said growth factor is selected from the group consisting of platelet-derived growth factor, fibroblast growth factor 2, and nerve growth factor.

6. The method of claim 1, wherein said neuronal differentiation-inducing compound is an anti-oxidant.

7. The method of claim 6, wherein said anti-oxidant is selected from the group consisting of beta-mercaptoethanol, dimethylsulfoxide, butylated hydroxytoluene, butylated hydroxyanisole, ascorbic acid, dimethylfumarate, and n-acetylcysteine.

8. The method of claim 7, wherein said anti-oxidant is beta-mercaptoethanol.

9. The method of claim 7, wherein said anti-oxidant is dimethylsulfoxide.

10. The method of claim 7, wherein said anti-oxidant is dimethylsulfoxide and butylated hydroxyanisole.

11. A method of producing an isolated neuronal cell, said method comprising isolating a marrow stromal cell, contacting said marrow stromal cell with a neuronal differentiation-inducing compound wherein said compound induces said isolated marrow stromal cell to differentiate into an isolated neuronal cell, thereby producing said isolated neuronal cell.

12. A method of treating a human patient having a disease, disorder or condition of the central nervous system, said method comprising obtaining a bone marrow sample from a human donor, isolating stromal cells from said bone marrow sample, inducing said stromal cells to differentiate into isolated neuronal cells, and administering said isolated neuronal cells to the central nervous system of said human patient, wherein the presence of said isolated neuronal cells in said central nervous system of said human patient effects treatment of said disease, disorder or condition.

13. The method of claim 12, wherein said disease, disorder or condition of the central nervous system is selected from the group consisting of Alzheimer'"'"'s disease, Parkinson'"'"'s disease, Huntington'"'"'s disease, amyotrophic lateral sclerosis, a tumor, a trauma, elderly dementia, Tay-Sach'"'"'s disease, Sandhoffs disease, Hurler'"'"'s syndrome, Krabbe'"'"'s disease, birth-induced traumatic central nervous system injury, epilepsy, multiple sclerosis, trauma, tumor, stroke, and spinal cord injury.

14. The method of claim 12, wherein prior to administering said isolated neuronal cells, said isolated neuronal cells are transfected with an isolated nucleic acid encoding a therapeutic protein, wherein when said protein is expressed in said cells said protein serves to effect treatment of said disease, disorder or condition.

15. The method of claim 14, wherein said isolated nucleic acid encodes a therapeutic protein selected from the group consisting of a cytokine, a chemokine, a neurotrophin, another trophic protein, a growth factor, an antibody, and a glioma toxic protein.

16. A method of treating a human patient in need of neuronal cells, said method comprising obtaining marrow stromal cells from a human patient, propagating said marrow stromal cells in culture under conditions that induce their differentiation into neuronal cells, transplanting said neuronal cells into said human patient in need of said neuronal cells, thereby treating said human patient in need of neuronal cells.

17. An isolated neuronal cell made by a method of inducing differentiation of an isolated marrow stromal cell, said method comprising contacting said isolated marrow stromal cell with at least one neuronal differentiation-inducing compound, thereby inducing differentiation of said isolated marrow stromal cell into said neuronal cell.

18. The cell of claim 17, wherein said cell is a human cell.

19. An isolated neuronal cell made by a method of inducing differentiation of an isolated marrow stromal cell, said method comprising contacting said isolated marrow stromal cell with at least one neuronal differentiation-inducing compound, thereby inducing differentiation of said isolated marrow stromal cell in said neuronal cell, wherein said neuronal cell is further transfected with an isolated nucleic acid encoding a therapeutic protein, and further wherein when said protein is expressed in said cell said protein serves to effect treatment of a disease, disorder, or condition of the central nervous system.

20. The cell of claim 19, wherein said isolated nucleic acid encodes a protein selected from the group consisting of a cytokine, a chemokine, a neurotrophin, another trophic protein, a growth factor, an antibody, and a glioma toxic protein.

21. The cell of claim 19, wherein said cell is a human cell.

22. An isolated neuronal cell made by a method of producing an isolated neuronal cell, said method comprising isolating a marrow stromal cell, contacting said marrow stromal cell with a neuronal differentiation inducing compound, wherein said compound induces said isolated marrow stromal cell to differentiate into said isolated neuronal cell, thereby producing said isolated neuronal cell.

23. The cell of claim 22, wherein said cell is a human cell.

24. An isolated neuronal cell made by a method of producing an isolated neuronal cell, said method comprising isolating a marrow stromal cell, contacting said marrow stromal cell with a neuronal differentiation inducing compound, wherein said compound induces said isolated marrow stromal cell to differentiate into said isolated neuronal cell, thereby producing said isolated neuronal cell, wherein said neuronal cell is further transfected with an isolated nucleic acid encoding a therapeutic protein, and further wherein when said protein is expressed in said cell, said protein serves to effect treatment of a disease, disorder, or condition of the central nervous system.

25. The cell of claim 24, wherein said isolated nucleic acid encodes a protein selected from the group consisting of a cytokine, a chemokine, a neurotrophin, another trophic protein, a growth factor, an antibody, and a glioma toxic protein.

26. The cell of claim 25, wherein said cell is a human cell.