Primary Rat Astroglial Culture in Serum-Free Supplemented Media and Effect of a Bovine Brain Factor

Abstract

A serum-free supplemented medium has been developed that supports growth of astroglial cells in primary culture from newborn rat brain hemispheres. This medium consists of a basal medium (Waymouth MD 705/1) supplemented with insulin (I), transferrin (T) and fatty-acid-free bovine serum albumin (BSA). In basal medium without serum only a few cells survived after 15 days. In the presence of I and T the number of cells increased. The same results was observed with the addition of a purified fraction from bovine brain extract (astroglial growth factor: AGF). Addition of AGF to the IT medium stimulated the proliferation of the cells and elicited an increase of the level of S-100 protein and of the activity of glutamine synthetase. The complete defined medium (with I, T and BSA) supported the growth of astroglial cells. After 14 days nearly all cells express glial fibrillary acidic (GFA) protein. These cells could be maintained in the defined medium over a period of 4 weeks.