In vitro differentiation of adult stem cells
First Claim
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1. A method of differentiating an adult bone marrow stromal cell into a desired cell type, said method comprising co-culturing said marrow stromal cell with another cell having said desired cell type, wherein said marrow stromal cell acquires the phenotype of said another cell.
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Abstract
The present invention includes a method for differentiating marrow stromal cells (MSCs) in vitro. The present invention also includes a method for improving the recovery rate of a mammal afflicted with a neuronal injury. The present invention also includes a method for enhancing the differentiation of marrow stromal cells to hypertrophic chondrocytes.
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25 Claims
- 1. A method of differentiating an adult bone marrow stromal cell into a desired cell type, said method comprising co-culturing said marrow stromal cell with another cell having said desired cell type, wherein said marrow stromal cell acquires the phenotype of said another cell.
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16. A method for improving recovery of a mammal from a neuronal injury, said method comprising culturing bone marrow stromal cells in a neurigenic medium and implanting said marrow stromal cells so cultured into the site of neuronal injury at least seven days post injury.
- 19. A method of enhancing production of hypertrophic chondrocyte cells, said method comprising co-culturing marrow stromal cells with chondrocytes in a hyperchondrogenic medium, said medium comprising from about 10 nanomolar to about 100 nanomolar beta glycerol phosphate.
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21. A hyperchondrogenic medium comprising about 20 nanomolar glycerol phosphate.
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22. A method of inducing a bone marrow stromal cell to produce a protein, said method comprising:
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(a) transfecting the bone marrow stromal cell with a DNA construct comprising;
a splice site;
a promoter for the expression of said protein;
a marker gene;
an internal ribosomal site (IRES);
an antibiotic resistance gene; and
another splice site;
(b) co-culturing said transfected bone marrow stromal cell with another damaged cell type obtained from an animal source;
(c) isolating those transfected bone marrow stromal cells that express both said marker gene and said antibiotic resistance gene; and
(d) optionally excising said DNA construct from said bone marrow stromal cell by incubation with recombinant Cre protein. - View Dependent Claims (23, 24)
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25. A method of inducing a bone marrow stromal cell to produce insulin, said method comprising:
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(a) transfecting the bone marrow stromal cell with a DNA construct comprising;
a lox site;
a promoter for the expression of insulin;
a green fluorescent protein gene;
an internal ribosomal site (IRES);
a neomycin resistance gene; and
another lox site;
(b) co-culturing said transfected bone marrow stromal cell with a heat-shocked beta cell obtained from an animal source;
(c) isolating those transfected bone marrow stromal cells that express both said green fluorescent protein gene and said neomycin resistance gene; and
(d) optionally excising said DNA construct from said bone marrow stromal cell by incubation with recombinant Cre protein.
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Specification