A procedure for long-term culture of oligodendrocytes

A procedure for long-term culture of oligodendrocytes is described, the starting material being 20-day-old primary mixed cultures of newborn rat brain. Cells were first incubated in a serum-free medium for 48 h before they were subcultured on poly-L-lysine coated plastic dishes. After this treatment, the oligodendrocytes developed well in Waymouth medium containing 10% (v/v) calf serum, while most of the astrocytes died. At 13 days in subculture more than 90% of the cells were identified as oligodendrocytes; the criteria for oligodendrocytes were based on their immunoreactivity to antisera against W1 Wolfgram protein, myelin basic proteins and the synthetic C-terminal hexapeptide of the major myelin proteolipid. At 13 and 19 days astrocytes were present, 7% and 20% respectively. The culture system described here may be useful to study the biochemical and immunological aspects of the oligodendrocytes.