Long-term Culture of Oligodendrocytes Isolated from Rat Corpus Callosum by Percoll Density Gradient

Oligodendrocytes isolated from the corpus callosum of four-week-old rats by trypsimization and Percoll density gradient centrifugation were cultured on poly-1-lysine coated coverslips. Some cells extended short processes within 24 hours (h), and at that time up to 95% of the cells showed surface binding of rabbit antiserum to galactocerebroside (anti-GalC) as demonstrated by indirect immunofluorescence. Oligodendrocytes survived up to two months in culture, extending processes with membranous elaborations. Exposure of living oligodendrocytes to varying dilutions of rabbit anti-GalC serum in the presence of complement produced cytotoxicity which was directly proportional to the concentration of antiserum and duration of exposure, as assessed by a nigrosin dye exclusion test. This system of isolating and culturing rat oligodendrocytes will permit further developmental and immunologic studies related to demyelinating diseases.