Chemically defined medium for rat astroglial cells in primary culture

We have developed a serum-free defined medium that supports the growth in primary culture of rat astroglial cells. Cells dissociated from cerebral hemispheres of newborn rats were maintained for 4 days in a basal medium (Waymouth's medium) containing 10% fetal calf serum, which was substituted by a serum-free medium. The basal medium was supplemented with insulin (5 μg/ml) and fatty acid free bovine serum albumin (0.5 mg/ml). Under these conditions the cells proliferate as estimated by cell counts and DNA content; however, growth was less than in Waymouth's medium supplemented with 10% fetal calf serum. In contrast, a very similar morphology was observed between cultures grown in the serum-free or serum-containing media. The serum-free medium allows some maturation of the astroglial cells as shown by the presence of glial fibrillary acidic (GFA) protein, S-100 protein and glutamine synthetase (GS) activity. The astroglial cells can survive and grow in this chemically defined medium for up to 5 weeks. The ability to culture astroglial cells in such a minimal defined medium should facilitate investigations concerning the effects of growth factors on their proliferation and maturation.