The differentiation of oligodendrocytes in a serum-free hormone-supplemented medium

Primary mixed cultures of trypsin-dissociated fetal and newborn rat brain and spinal cord have been grown in a serum-free medium. This medium, containing insulin, selenium, transferrin and triiodothyronine, was optimized for oligodendrocyte survival by determining the number of cells which expressed surface galactocerebroside. Comparison of cultures in serum-containing and serum-free media revealed that galactocerebroside positive (GalC+) oligodendrocytes could be detected earlier in the absence of serum. This early differentiation occurred in the absence of the added hormones and nutrients, whose main function appeared to be to prolong survival of the cells. The effect of serum on the differentiation of oligodendrocytes was studied by comparing the expression of surface GalC in media containing 2.5% or 10% fetal calf serum. At a given time a much greater number of GalC+ oligodendrocytes could be detected at the lower serum concentration. However, when cultures were transferred from 10% serum to serum-free medium (or 1% serum) large numbers of GalC+ oligodendrocytes subsequently appeared, showing that precursors were present in the high-serum medium, but that they were unable to differentiate. Possible explanations of the effect of serum on oligodendrocyte differentiation are discussed.