Human CNS cell lines and methods of use therefor

Abstract

Conditionally-immortalized human CNS progenitor cell lines are provided. Such cell lines, which may be clonal, may be used to generate neurons and/or astrocytes. Such cell lines and/or differentiated cells may be used for the development of therapeutic agents to prevent and treat a variety of CNS-related diseases. Such cell lines and/or differentiated cells may also be used in assays and for the general study of CNS cell development, death and abnormalities.

1 - 11 . (Canceled)  
     
     
         12 . A method for introducing a CNS cell into a mammal, comprising administering to a mammal a cell produced by a method comprising: 
 (a) plating human CNS progenitor cells on a surface that permits proliferation, said surface being tissue culture plastic or a surface treated with fibronectin;    (b) adding serum-free growth medium to the cells;    (c) allowing the CNS progenitor cells to proliferate in the serum-free medium;    (d) transfecting the cells with DNA encoding a selectable marker and regulatable growth-promoting gene, wherein the growth-promoting gene is selected from the group consisting of SV40 large T antigen, v-myc, N-myc, c-myc, p53, polyoma large T antigen, E1a adenovirus and E7 protein of human papilloma virus;    (e) passaging the transfected cells onto a substrate; and    (f) adding serum-free growth medium containing one or more proliferation-enhancing factors to the transfected cells, wherein said proliferation-enhancing factors are selected from the group consisting of FGF-2, PDGF, EGF, medium conditioned by perpetualized adult rat hippocampal progenitor cells, and a combination thereof, therefrom producing a conditionally-immortalized human CNS progenitor cell.    
     
     
         13 . A method for introducing a CNS cell into a mammal, comprising administering to a mammal a conditionally-immortalized clonal human CNS progenitor cell capable of differentiation into neurons and astrocytes.  
     
     
         14 . A method for treating a patient, comprising administering to a patient a cell produced by a method comprising: 
 (a) plating human CNS progenitor cells on a surface that permits proliferation, said surface being tissue culture plastic or a surface treated with fibronectin;    (b) adding serum-free growth medium to the cells;    (c) allowing the CNS progenitor cells to proliferate in the serum-free medium;    (d) transfecting the cells with DNA encoding a selectable marker and regulatable growth-promoting gene, wherein the growth-promoting gene is selected from the group consisting of SV40 large T antigen, v-myc, N-myc, c-myc, p53, polyoma large T antigen, E1a adenovirus and E7 protein of human papilloma virus;    (e) passaging the transfected cells onto a substrate; and    (f) adding serum-free growth medium containing one or more proliferation-enhancing factors to the transfected cells, wherein said proliferation-enhancing factors are selected from the group consisting of FGF-2, PDGF, EGF, medium conditioned by perpetualized adult rat hippocampal progenitor cells, and a combination thereof, therefrom producing a conditionally-immortalized human CNS progenitor cell.    
     
     
         15 . A method for treating a patient, comprising administering to a mammal a conditionally-immortalized clonal human CNS progenitor cell capable of differentiation into neurons and astrocytes.  
     
     
         16 . A method according to  claim 15  wherein the patient is afflicted with a pathological condition where neurons have degenerated.  
     
     
         17 . A method according to  claim 16  wherein the pathological condition is selected from the group consisting of Alzheimer's disease, Parkinson's disease, amylotrophic lateral sclerosis, stroke and traumatic head injury.  
     
     
         18 - 32 . (Canceled)