Primary glial cells and brain fibroblasts: interactions in culture

Primary glial-enriched cultures were prepared from newborn mouse cerebral hemispheres. The cultures were grown in Dulbecco's Modified Eagle Medium in which L-valine was substituted with D-valine; this medium selectively inhibits the growth of fibroblasts. Using glutamine synthetase and glial fibrillary acidic protein as immunocytochemical markers, cultures in D-valine medium were characterized as being over 80% astrocytic. However, these cultures exhibited a suppressed growth rate and lagged behind in their differentiation as assessed biochemically using DNA content and glutamine synthetase activity as markers for growth and differentiation. Growth was restored when D-valine cultures were grown in medium containing conditioned medium derived from brain fibroblast cultures when grown on matrix or killed substrata derived from brain fibroblast cultures. This in vitro approach offers the possibility of purifying factors and developing immunological probes to investigate the possible role of brain fibroblasts in influencing glial cell function.