Patent ID: 6713247
Filing Date: 2004-03-30
Classification: A61K,A61P,C12N

Abstract:
A method for producing a conditionally-immortalized human CNS cell, 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 encodes a protein 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; and therefrom producing a conditionally-immortalized human CNS progenitor cell that differentiates only upon suppression of the growth-promoting protein.