Transfection of transformed shiverer mouse glial cell lines

In the course of studies on glial cell differentiation in the mouse mutant shiverer, we have established by infection with a temperature-sensitive retrovirus encoding the SV40 T antigen a few glial cell lines that divide rapidly at 33 degrees C, the permissive temperature, and whose phenotypes at the nonpermissive temperature (39 degrees C) resemble either astrocytes or oligodendrocytes. One astrocyte-like clone (clone SF11) expresses glial fibrillary acidic protein at 33 degrees C and expresses this marker strongly at 39 degrees C as well. One oligodendrocyte-like clone (DM5) that we have succeeded in maintaining at 33 degrees C can be induced to express galactocerebroside and 2',3'-cyclic nucleotide 3'-phosphodiesterase at the elevated temperature but never expresses the more differentiated markers of the myelination state, such as the myelin proteolipid protein or the myelin-associated glycoprotein. Clone DM5, as is the case with other oligodendrocyte clones we have prepared by the method described here, is quite fragile and survives for only several (5-6) days at the higher temperature. Both clonal lines can serve as host cells for expressible cDNAs introduced by transfection, such as the neurofilament protein, NF-M, and the small myelin basic protein.