A rapid procedure for the preparation of oligodendrocyte-enriched cultures from rat spinal cord.
Spinal cords and cerebra from 7-day-old rat pups were compared as tissue sources for the isolation of oligodendrocytes and for studies on the development of these cells in culture. After 1 day in culture the serum-containing medium was replaced by a chemically-defined medium, which contained a cocktail of hormones that stimulated oligodendrocyte development. The cultures were characterized with various immunocytochemical markers; monoclonal A2B5 for bipotential glial progenitor cells, anti-galactocerebroside (GC) serum for oligodendrocytes, and anti-glial fibrillary acidic protein (GFAP) serum for astrocytes. The number of positive cells was counted and expressed as a percentage of total cells. At 1 day in culture the cell cultures from spinal cord contained 30% GC+ cells, increasing to 90% after 7 days in culture. In cultures derived from cerebra the percentage of GC+ cells was always lower than in cultures from spinal cord. In cerebral cultures GFAP+ cells increased from 15% at 1 day in culture to 30% at 7 days in culture, whereas it remained low in spinal cord cultures. The activity of oligodendroglial marker enzyme 2',3'-cyclic-nucleotide 3'-phosphodiesterase was followed during development in culture. The specific activity increased rapidly in both types of culture but was more than threefold higher in cultures derived from spinal cord. This procedure yields, within one week and without subculture, primary glial cultures from rat spinal cord, that are highly enriched in oligodendrocytes (greater than or equal to 90%; 3.10(5) oligodendrocytes per rat pup).
van der Pal
R H
RH
Laboratory of Veterinary Biochemistry, University of Utrecht, The Netherlands.
Vos
J P
JP
van Golde
L M
LM
Lopes-Cardozo
M
M
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000032:analysis; D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002478:Cells, Cultured; D003712:Dendrites / Q000201:enzymology; D005260:Female; D005455:Fluorescent Antibody Technique; D005699:Galactosylceramides / Q000032:analysis; D011247:Pregnancy; D051381:Rats; D011919:Rats, Inbred Strains; D013116:Spinal Cord / Q000166:cytology* / Q000378:metabolism
D005699:Galactosylceramides; C002454:galactocerebroside; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases