Multi-lineage directed induction of bone marrow stromal cell differentiation
First Claim
1. A method of inducing differentiation of an isolated marrow stromal cell into an endodermal cell, said method comprising contacting said isolated marrow stromal cell with at least one endodermal/neuronal precursor differentiation-inducing compound, thereby inducing differentiation of said isolated marrow stromal cell into a endodermal/neuronal precursor cell and contacting said endodermal/neuronal precursor cell with at least one endodermal differentiation-inducing compound thereby inducing an endodermal cell.
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Abstract
Methods of inducing differentiation of mammalian bone marrow stromal cells into cells of multiple embryonic lineages by contacting marrow stromal cells with precursor differentiation-inducing compounds followed by contacting the partially differentiated precursor cells with specific cell type differentiation-inducing compounds. In one embodiment, the MSC derived precursor cell cultures comprise cells, at least some of which simultaneously express markers that are characteristic of endodermal and ectodermal cell types. In another embodiment, the differentiated cells are insulin-secreting pancreatic islet cells. Precursor 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. Endodermal cell differentiation-inducing compounds of the invention include but are not limited to anti-oxidants and growth factors including basic fibroblast growth factor. Once induced to differentiate into a particular cell type, the cells can be used for cell therapy, gene therapy, or both, for treatment of diseases, disorders, or conditions associated with tissues of multiple embryonic origins.
87 Citations
50 Claims
- 1. A method of inducing differentiation of an isolated marrow stromal cell into an endodermal cell, said method comprising contacting said isolated marrow stromal cell with at least one endodermal/neuronal precursor differentiation-inducing compound, thereby inducing differentiation of said isolated marrow stromal cell into a endodermal/neuronal precursor cell and contacting said endodermal/neuronal precursor cell with at least one endodermal differentiation-inducing compound thereby inducing an endodermal cell.
- 20. A method of producing an isolated endodermal cell, said method comprising isolating a marrow stromal cell, contacting said isolated marrow stromal cell with at least one endodermal/neuronal precursor differentiation-inducing compound, thereby inducing differentiation of said isolated marrow stromal cell into a endodermal/neuronal precursor cell and contacting said endodermal/neuronal precursor cell with an endodermal differentiation-inducing compound thereby inducing an endodermal cell.
- 22. A method of treating a human patient having a disease, disorder or condition of an endodermal tissue, 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 selected isolated endodermal cells, and administering said selected isolated endodermal cells to the body of said human patient, wherein the presence of said selected isolated neuronal cells in said body of said human patient effects treatment of said disease, disorder or condition.
- 26. A method of treating a human patient in need of endodermal 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 selected endodermal cells, transplanting said selected endodermal cells into said human patient in need of said selected endodermal cells, thereby treating said human patient in need of endodermal cells.
- 28. An isolated endodermal 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 endodermal/neuronal precursor differentiation-inducing compound, thereby inducing differentiation of said isolated marrow stromal cell into a endodermal/neuronal precursor cell and contacting said endodermal/neuronal precursor cell with an endodermal differentiation-inducing compound thereby inducing an endodermal cell.
- 31. An isolated endodermal 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 endodermal/neuronal precursor differentiation-inducing compound, thereby inducing differentiation of said isolated marrow stromal cell into a endodermal/neuronal precursor cell and contacting said endodermal/neuronal precursor cell with an endodermal differentiation-inducing compound thereby inducing an endodermal cell wherein said endodermal cell is further transfected with an isolated nucleic acid encoding a therapeutic protein or peptide, and further wherein when said protein or peptide is expressed in said cell said protein or peptide serves to effect treatment of a disease, disorder, or condition associated with a tissue of endodermal origin.
- 34. An isolated endodermal cell made by a method of producing an isolated endodermal cell, said method comprising isolating a marrow stromal cell, contacting said isolated marrow stromal cell with at least one endodermal/neuronal precursor differentiation-inducing compound, thereby inducing differentiation of said isolated marrow stromal cell into a endodermal/neuronal precursor cell and contacting said endodermal/neuronal precursor cell with an endodermal differentiation-inducing compound thereby inducing said endodermal cell
- 37. An isolated endodermal cell made by a method of producing an isolated endodermal cell, said method comprising isolating a marrow stromal cell, contacting said isolated marrow stromal cell with at least one endodermal/neuronal precursor differentiation-inducing compound, thereby inducing differentiation of said isolated marrow stromal cell into a endodermal/neuronal precursor cell and contacting said endodermal/neuronal precursor cell with an endodermal differentiation-inducing compound thereby inducing said endodermal cell wherein said endodermal cell is further transfected with an isolated nucleic acid encoding a therapeutic protein or peptide, and further wherein when said protein or peptide is expressed in said cell, said protein or peptide serves to effect treatment of a disease, disorder, or condition associated with a tissue of endodermal origin.
- 41. An MSC derived cell culture that comprises cells, at least some of which simultaneously express polypeptide or mRNA markers that are characteristic of at least endodermal and ectodermal cell types.
- 46. A method of producing an isolated endodermal/neuronal precursor cell, said method comprising isolating a marrow stromal cell culturing the marrow stromal cell under conditions suitable to produce an endodermal/neural precursor culture that comprises cells at least some of which simultaneously express polypeptide or mRNA markers that are characteristic of at least endodermal and ectodermal cell types.
Specification