GENERATION AND ANALYSIS OF NORMAL AND SHIVERER TEMPERATURE-SENSITIVE IMMORTALIZED CELL-LINES EXHIBITING PHENOTYPIC CHARACTERISTICS OF OLIGODENDROCYTES AT SEVERAL STAGES OF DIFFERENTIATION

Normal glial cells immortalized at specific developmental stages would be useful tools with which to study glial cell differentiation in vit ro. Similarly, immortalized glial cell lines derived from known neurol ogical mutants with identified developmental, molecular genetic defect s would also be useful for the in vitro examination of the effects of the mutation on glial cell function. In this report we describe the im mortalization of 19 separate oligodendroglial cell lines, 10 derived f rom normal mice and 9 derived from the neurological mutant shiverer, w hich is missing a large segment of the myelin basic protein gene. Enri ched oligodendrocyte cultures prepared at 7 days in vitro, a time when the majority of the cells were oligodendrocyte precursors undergoing transition into mature oligodendrocytes, were immortalized with pZIPSV tsA58, which carries a temperature-sensitive immortalizing oncogene. A ll of the immortalized cell lines grew rapidly at the permissive tempe rature of 34 degrees C and exhibited a dramatic decrease in growth rat e at the nonpermissive temperature of 39 degrees C. The cell lines wer e characterized by immunocytochemistry and Northern blot analysis for a number of glial cell markers, and the phenotype of all the lines wer e consistent with their being in the oligodendroglial cell lineage. Th e phenotypes of the cell lines varied significantly with representativ es of oligodendrocyte precursors, and immature and mature oligodendroc ytes being present within the population of immortalized lines. All th e cell lines appeared to be clonal based upon Southern blot analysis a nd all have been passaged at least 40 times with retention of stable p henotype.