Chemically defined medium for rat astroglial cells in primary culture

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Abstract
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.