Transdifferentiation of glial cells

Abstract

A process for generating multipotent cells from glial cells using in vitro techniques to dedifferentiate fetal or adult mammalian glial cells into multipotent cells. The multipotent cells may further be differentiated into particular types of nervous system cells, including neurons, astrocytes, and oligodendrocytes. A small sample of astrocytes is used to establish an in vitro culture of cells that is expanded and processed to yield multipotent cells that may be used directly or be differentiated to yield neurons and/or oligodendrocytes and/or astrocytes. The invention includes implanting the generated cells into patients. The invention includes a step of exposing the cells to a growth factor.

1 . A method of delivering a therapy to a human patient the method comprising providing a group of cells and introducing the group of cells into a therapy site in the patient, wherein the group of cells comprise neurons and/or oligodendrocytes cultured in vitro from a confluent layer of autologous astrocytes.  
   
   
       2 . The method of  claim 1  comprising the step of choosing the therapy site from the group consisting of the central nervous system, the brain, the cranium, the spine, the peripheral nervous system, the region interior to the skull and the ganglia.  
   
   
       3 . The method of  claim 1  wherein the group of cells are cultured by treating the autologous astrocytes with a medium comprising bFGF.  
   
   
       4 . The method of  claim 3  wherein the concentration of the bFGF is from about 0.05 to about 1,000 ng per ml.  
   
   
       5 . The method of  claim 3  wherein the autologous astrocytes are treated with the medium comprising a FGF for at least 10 days.  
   
   
       6 . The method of  claim 3  wherein the medium further comprises heparin.  
   
   
       7 . The method of  claim 1  further comprising dissociating and plating the group of cells prior to introducing the group of cells into the therapy site.  
   
   
       8 . The method of  claim 7  wherein the group of cells are plated onto a substrate that comprises a member of the group consisting poly-L-lysine, polyornithine, and extracellular matrix.  
   
   
       9 . The method of  claim 1  wherein the autologous astrocytes are derived from mammalian neural stem cells.  
   
   
       10 . The method of  claim 1  wherein the group of cells are cultured by maintaining the cells in a medium essentially free of bFGF for at least one day.  
   
   
       11 . The method of  claim 10  wherein the medium comprises DMEM.  
   
   
       12 . The method of  claim 10  wherein the medium comprises F12.  
   
   
       13 . The method of  claim 10  wherein the medium comprises FGF-8.  
   
   
       14 . The method of  claim 10  wherein the medium comprises a member of the group consisting of retinoic acid, dbcAMP, BDNF and GDNF.