PREPARATION OF VIABLE ASTROCYTES FROM THE DEVELOPING CEREBELLUM

A cell fraction enriched in viable astrocytes has been prepared from the developing rat cerebellum. The isolated astrocytes were identified by indirect immuno-flourescence using antiserum to glial fibrillry acidic protein (GFAP). In comparison with the low incidence (6%) of GFAP-positive cells in the unfractionated cell suspension from trypsinized cerebellar tissue, these accounted for half of the cells in the final preparation. This was achieved by treatment at 6 days of age with hydroxyurea (an inhibitor of DNA synthesis) which enriched the tissue in GFAP-positive astrocytes at day 8. At the same time, by selectively removing the heterogeneously sized replicating cells, hydroxyurea treatment resolution of a fraction enriched in GFAP-positive cells using unit gravity sedimentation. The isolated astrocytes differentiated in culture producing, by 3 days in vitro, a dense network of fine GFAP-containing processes. © 1979.