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Culture of rat cerebral oligodendrocytes in a serum-free, chemically defined medium

By J.W. Koper, M. Lopes-Cardozo, H.J. Romijn and L.M.G. van Golde


Oligodendrocytes were isolated from the cerebra of young rats (5-10 days old) by trypsinization of the\ud tissue followed by cell separation on Percoll gradients. The isolation was carried out in physiological,\ud isotonic media. The cell yield was 2-4 × 10⁶ cells per brain; the plating efficiency was ≥70%. Isolated\ud cells were seeded on poly-L-lysine-coated culture dishes and maintained in a serum-free, chemically\ud defined medium for at least 30 days. After 10 days in culture 67±10% of the surviving cells were\ud oligodendrocytes, as judged by immunocytochemical and morphological criteria, whereas most of the\ud other cells reacted positively with antiserum against glial fibrillary acidic protein. The expression of\ud typical oligodendrocyte markers (2':3'-cyclic-nucleotide 3'-phosphodiesterase, galactocerebrosides and\ud myelin basic protein) was greatly enhanced under these serum-free conditions as compared with cultures\ud in serum-containing medium. The antigenic markers (galactocerebrosides, myelin basic protein) were\ud absent in the freshly isolated cells but could be detected after 3 days in culture by immunocytochemistry.\ud The activity of 2':3'-cyclic-nucleotide 3'-phosphodiesterase increased from 75 nmol min⁻¹ mg⁻¹ protein\ud on day 4 to 400 nmol min⁻¹ mg⁻¹ protein on day 14 in culture

Topics: Diergeneeskunde, oligodendrocyte; primary culture; serum-free medium;, chemically defined medium; brain cells; rat brain, oligodendrocyte, primary culture, serum-free medium, chemically defined medium, brain cells, rat brain
Year: 1984
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