Cells exhibiting neuronal progenitor cell characteristics and methods of making them

Abstract

Disclosed are cells exhibiting neuronal progenitor cell characteristics, and methods of making them from marrow adherent stem cells by regulating cellular pathways in the marrow adherent stem cells that are associated with glial transdifferentiation of the marrow adherent stem cells.

1 . A method for making a cell for treatment of a disease of the central or peripheral nervous system, wherein said cell is obtained by:
 (a) obtaining a bone marrow sample;   (b) isolating cells from the bone marrow sample;   (c) incubating the cells in tissue culture;   (d) removing non-adherent cells from the culture to provide a culture of marrow adherent stem cells (MASCs);   (e) incubating the MASCs with a JAK/STAT inhibitor, wherein the MASCs are not transfected with a polynucleotide encoding the Notch intracellular domain; and   (f) isolating, from the culture, a cell that:
 (i) is mitotic, 
 (ii) expresses EfnB2, 
 (iii) expresses nestin, 
 (iv) does not express CD90, and 
 (v) does not express PDGF receptor beta. 
   
     
     
         2 . The method of  claim 1 , wherein the marrow adherent stem cells are selected from the group consisting of human marrow adherent stem cells, rat marrow adherent stem cells, mouse marrow adherent stem cells, primate marrow adherent stem cells, pig marrow adherent stem cells, cow marrow adherent stem cells, and sheep marrow adherent stem cells. 
     
     
         3 . The method of  claim 2 , wherein the marrow adherent stem cells are human marrow adherent stem cells. 
     
     
         4 . The method of  claim 1 , wherein the inhibitor of JAK/STAT signal transduction comprises an inhibitor of STAT1 or STAT3. 
     
     
         5 . The method of  claim 1 , wherein the JAK/STAT inhibitor is a polypeptide. 
     
     
         6 . The method of  claim 5 , wherein the incubation comprises transfection of the MASCs with a polynucleotide encoding the JAK/STAT inhibitor. 
     
     
         7 . The method of  claim 1 , wherein the JAK/STAT inhibitor is 4-(4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline. 
     
     
         8 . The method of  claim 1 , further comprising:
 administering, to a subject, one or more cells according to step (f).   
     
     
         9 . An isolated cell, wherein said cell is obtained by:
 (a) obtaining a bone marrow sample;   (b) isolating cells from the bone marrow sample;   (c) incubating the cells in tissue culture;   (d) removing non-adherent cells from the culture to provide a culture of marrow adherent stem cells (MASCs);   (e) incubating the MASCs with a JAK/STAT inhibitor, wherein the MASCs are not transfected with a polynucleotide encoding the Notch intracellular domain; and   (f) isolating, from the culture, a cell that:
 (i) is mitotic, 
 (ii) expresses EfnB2, 
 (iii) expresses nestin, 
 (iv) does not express CD90, and 
 (v) does not express PDGF receptor beta. 
   
     
     
         10 . The cell of  claim 9 , wherein the marrow adherent stem cells are selected from the group consisting of human marrow adherent stem cells, rat marrow adherent stem cells, mouse marrow adherent stem cells, primate marrow adherent stem cells, pig marrow adherent stem cells, cow marrow adherent stem cells, and sheep marrow adherent stem cells. 
     
     
         11 . The cell of  claim 10 , wherein the marrow adherent stem cells are human marrow adherent stem cells. 
     
     
         12 . The cell of  claim 9 , wherein the inhibitor of JAK/STAT signal transduction comprises an inhibitor of STAT1 or STAT3. 
     
     
         13 . The cell of  claim 9 , wherein the JAK/STAT inhibitor is a polypeptide. 
     
     
         14 . The cell of  claim 13 , wherein the incubation comprises transfection of the MASCs with a polynucleotide encoding the JAK/STAT inhibitor. 
     
     
         15 . The cell of  claim 9 , wherein the JAK/STAT inhibitor is 4-(4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline. 
     
     
         16 . A method comprising administering the cell of  claim 9  to a subject. 
     
     
         17 . An isolated population of cells descended from marrow adherent stem cells, wherein the population contains a plurality of cells that:
 (i) are mitotic,   (ii) express EfnB2,   (iii) express nestin,   (iv) do not express CD90, and   (v) do not express PDGF receptor beta;   and further wherein said marrow adherent stem cells are not transfected with a polynucleotide encoding the Notch intracellular domain.   
     
     
         18 . A method comprising administering, to a subject, a cell population according to  claim 17 .