Primary culture and cryopreservation of mouse astrocytes under serum-free conditions.
The methods of primary culture and cryopreservation of mouse astrocytes under serum-free conditions were examined. Cerebra from newborn C3H/He mice were employed as the source of astrocytes. The cultured cells were able to grow in a serum-free, chemically defined medium containing transferrin, hydrocortisone, biotin, sodium selenite, insulin, fibroblast growth factor and epidermal growth factor. After the culture was maintained in the medium for 3 weeks, purity was assessed using immunofluorescence staining. The great majority of the cells (greater than 98%) contained glial fibrillary acidic protein and S-100 protein which are cell markers of astrocytes. To cryopreserve the enriched astrocytes under serum-free conditions, various cryoprotectants were examined. The combination of 10% dimethylsulfoxide and 0.1% methylcellulose gave the highest survival rate. These methods of primary culture and cryopreservation will be useful in physiological and biochemical studies which require mouse astrocytes.
Yoshida
T
T
Institute for Fermentation, Osaka, Japan.
Takeuchi
M
M
D000818:Animals; D001253:Astrocytes / Q000166:cytology*; D001769:Blood; D001921:Brain / Q000166:cytology; D002448:Cell Adhesion; D002455:Cell Division; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation; D003470:Culture Media; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D051379:Mice; D008809:Mice, Inbred C3H
D003470:Culture Media; D004121:Dimethyl Sulfoxide