Rapidly proliferating glial cells isolated from adult mouse brain have a differentiative capacity in response to...

A glial cell line designated as B2 was generated from primary cultures of oligodendrocytes/astrocytes isolated from an adult BALB/c mouse brain and maintained for over 1 year. Phenotypic characteristics of B2 cells were investigated by immunolabeling with cell type-specific markers for oligodendrocytes (O4 and galactocerebroside (GalC)), astrocytes (glial fibrillary acidic protein (GFAP)), and immature neuroectodermal cells (vimentin). When cultured in a serum-containing medium, B2 cells exhibited a bipolar or a tripolar process-bearing morphology and proliferated with a 24-28 h doubling time, without requirement of exogenous growth factors. Under this culture condition, vimentin was identified in all of the B2 cells, GFAP in 7%, and O4 and GalC in less than 1% of the cells. When cultured in a serum-free medium containing 1 mM dibutyryl cyclic AMP (dbcAMP), B2 cells extended longer processes and 45% of the cells expressed cell type-specific markers for oligodendrocytes or astrocytes. GFAP was identified in 29% of B2 cells, O4 in 16%, and GalC in 6% of the cells, although, neither O4+GFAP+ nor GalC+GFAP+ cells were observed. B2 cells proliferated in response to phorbol 12-myristate 13-acetate (PMA), basic fibroblast growth factor (bFGF), insulin, insulin-like growth factor (IGF-I) and platelet-derived growth factor (PDGF), but not to dbcAMP, forskolin (FK), or retinoic acid (RA). These results indicate that B2 cells are distinct from typical oligodendrocytes and astrocytes with respect to their great proliferative potential, and suggest that B2 cells, with a capacity to differentiate into oligodendrocytes and astrocytes in response to cyclic AMP, may represent a population of glial precursor cells in the adult mouse central nervous system (CNS).