Cultivation of immature astrocytes of mouse cerebellum in a serum-free, hormonally defined medium. Appearance of the mature astrocyte phenotype after addition of serum.
A hormonally defined culture medium is described which supports survival and proliferation of astroglia from primary cultures of early postnatal mouse cerebellum. This medium consists of bovine serum albumin, insulin, transferrin, selenium, hyaluronic acid, protease inhibitor aprotinin, and epidermal growth factor. Trypsin-dissociated single cerebellar cell suspensions are plated in this medium on poly-l-coated glass coverslips and maintained for two weeks before subcultivation. After subcultivation into defined medium more than 99% of all cells are vimentin-positive and fibronectin- and almost completely glial fibrillary acid (GFA) protein-negative, indicating that these cells are less mature astrocytes. After replacement of defined medium by culture medium containing 10% horse serum, expression of GFA protein is detectable in addition to vimentin by indirect immunofluorescence.
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000502:physiology*; D001769:Blood; D002455:Cell Division; D002478:Cells, Cultured; D002531:Cerebellum / Q000502:physiology*; D003470:Culture Media; D051379:Mice; D008810:Mice, Inbred C57BL
D003470:Culture Media