Generation, Characterization and Isolation of Neuroepithelial Stem Cells and Lineage Restricted Intermediate Precursor

Abstract

Multipotent neuroepithelial stem cells and lineage-restricted oligodendrocyte-astrocyte precursor cells are described. The neuroepithelial stem cells are capable of self-renewal and of differentiation into neurons, astrocytes, and oligodendrocytes. The oligodendrocyte-astrocyte precursor cells are derived from neuroepithelial stem cells, are capable of self-renewal, and can differentiate into oligodendrocytes and astrocytes, but not neurons. Methods of generating, isolating, and culturing such neuroepithelial stem cells and oligodendrocyte-astrocyte precursor cells are also disclosed.

1 . An isolated population of mammalian CNS neuroepithelial stem cells capable of self-renewal, wherein said cells express nestin, but do not express polysialated neural cell adhesion molecule, glial fibrillary acidic protein, sulfatide, neurofilament, choline acetyl transferase, intermediate filament, ganglioside, or galactocerebroside and wherein said cells differentiate into neurons and glial of the central nervous system. 
   
   
       2 . The isolated population of mammalian CNS neuroepithelial stem cells of  claim 1  in a feeder-cell-independent adherent culture. 
   
   
       3 . The isolated population of mammalian CNS neuroepithelial stem cells of  claim 1  which require fibroblast growth factor and chick embryo extract to self-renew and which differentiate in the absence of chick embryo extract. 
   
   
       4 . A population of central nervous system glia and neurons, said population differentiated from the isolated population of mammalian CNS neuroepithelial stem cells of  claim 1 . 
   
   
       5 . An isolated population of mammalian CNS glial-restricted precursor cells capable of self-renewal, wherein said cells express nestin and ganglioside, but do not express glial fibrillary acidic protein, sulfatide, or galactocerebroside and wherein said cells differentiate into oligodendrocytes, type 1 astrocytes and type 2 astrocytes, but do not differentiate into neuronal cells. 
   
   
       6 . A population of oligodendrocytes, type 1 astrocytes and type 2 astrocytes differentiated from the isolated population of mammalian CNS glial-restricted precursor cells of  claim 5 .