Immortalization of different precursors of glial cells with a targeted and temperature-sensitive oncogene

Different types of glial precursor cell lines were obtained after stable transfection of brain cells with the pJC-SVLTtsA vector carrying the tsA58 simian virus 40 large T (SVLT) gene driven by the promoter of a gliotropic strain of JC papovavirus. The immortalized cells were conditional for growth: they expressed the SVLT antigen and proliferated at 34 degrees C, but their growth was either reduced or arrested when they were shifted to 39 degrees C. The differentiation characteristics of four representative lines were more extensively studied. The CR15 and CM8 lines displayed properties of bipotential glial progenitors: they were able to express oligodendrocyte markers at both temperatures, but could differentiate into astrocytes only at 39 degrees C. In contrast, the CR19 and CM3r lines corresponded to more committed oligodendrocyte precursors: they expressed various oligodendroglial markers but they could not synthesize the glial fibrillary acidic protein. More particularly, the CM3r mouse cells displayed a typical oligodendrocyte progenitor morphology and expressed the proteolipid protein mRNA.