Trans-Differentiation And Re-Differentiation Of Somatic Cells And Production Of Cells For Cell Therapies

Abstract

The invention provides a method for effecting the trans-differentiation of a somatic cell, i.e., the conversion of a somatic cell of one cell type into a somatic cell of a different cell type. The method is practiced by culturing a somatic cell in the presence of at least one agent selected from the group consisting of (a) cytoskeletal inhibitors and (b) inhibitors of acetylation, and (c) inhibitors of methylation, and also culturing the cell in the presence of agents or conditions that induce differentiation to a different cell type. The method is useful for producing histocompatible cells for cell therapy.

1 . A method for effecting trans-differentiation of a somatic cell comprising
 (a) culturing a somatic cell in the presence of at least one agent selected from the group consisting of (a) cytoskeletal inhibitors and (b) inhibitors of acetylation, and (c) inhibitors of methylation;   (b) culturing the cell in the presence of agents or conditions that induce differentiation to a cell type different than that of said somatic cell; and   (c) allowing the cell to trans-differentiate to said different cell type.   
     
     
         2 . The method of  claim 1 , wherein the somatic cell is a mammalian cell selected from the group consisting of fibroblasts, B cells, T cells, dendritic cells, keratinocytes, adipose cells, epithelial cells, epidermal cells, chondrocytes, cumulus cells, neural cells, glial cells, astrocytes, cardiac cells, esophageal cells, muscle cells, melanocytes, hematopoietic cells, macrophages, monocytes, and mononuclear cells. 
     
     
         3 . The method of  claim 1 , wherein the somatic cell is a mammalian fibroblast. 
     
     
         4 . The method of  claim 1 , further comprising culturing the somatic cell in serum-free medium before or after performing step (a). 
     
     
         5 . The method of  claim 1 , wherein step (a) comprises culturing the somatic cell in the presence of at least one cytoskeletal inhibitor selected from the group consisting of microfilament inhibitors and microtubule inhibitors. 
     
     
         6 . The method of  claim 5 , wherein the at least one microfilament inhibitor is selected from the group consisting cytochalasin B, D, A, E; vimentin, latrunculin, and jasplakinolide. 
     
     
         7 . The method of  claim 5 , wherein the at least one microfilament inhibitor is cytochalasin B. 
     
     
         8 . The method of  claim 5 , wherein the at least one microtubule inhibitor is selected from the group consisting of colchicine, colcemid, nocodazole, and taxol. 
     
     
         9 . The method of  claim 1 , wherein the at least one inhibitor of acetylation is a deacetylase inhibitor. 
     
     
         10 . The method of  claim 1 , wherein the at least one inhibitor of methylation is a methylase inhibitor. 
     
     
         11 . The method of  claim 1 , further comprising culturing the somatic cell in serum-free medium after performing step (a). 
     
     
         12 . The method of  claim 1 , wherein the cell is cultured in the presence of agents or conditions that induce differentiation to a different cell type before or after performing step (a). 
     
     
         13 . The method of  claim 1 , wherein the cell is cultured in the presence of agents or conditions that induce differentiation to a cell of neuronal lineage. 
     
     
         14 . The method of  claim 13 , wherein cell is cultured in keratinocyte growth medium or neuro-progenitor growth medium before or after performing step (a). 
     
     
         15 . The method of  claim 13 , further comprising assaying to detect a marker of cells of neural lineage. 
     
     
         16 . A method for converting a non-neural somatic cell to become a cell having the phenotype of a cell of neural lineage, comprising:
 (a) culturing a non-neural somatic cell in the presence of at least one agent selected from the group consisting of (a) cytoskeletal inhibitors and (b) inhibitors of acetylation, and (c) inhibitors of methylation;   (b) culturing the cell in the presence of agents or conditions that induce differentiation to a cell of neuronal lineage; and   (c) allowing the cell to trans-differentiate to a cell having the phenotype of a cell of neural lineage.   
     
     
         17 . The method of  claim 16 , wherein the somatic cell is a fibroblast. 
     
     
         18 . The method of  claim 16 , wherein step (b) comprises cultured the cell in keratinocyte growth medium or neuro-progenitor growth medium. 
     
     
         19 . The method of  claim 18 , further comprising assaying to detect a marker of cells of neural lineage. 
     
     
         20 . A composition of cells having the phenotype of cells of neural lineage. prepared by the method of  claim 16 .