Astrocytes (McGill University)

Fetal brain cells were dissociated from the fetal brain tissue of 14-18-week-old human fetuses, using trypsin and deoxyribonuclease I. The cerebral tissue was minced in serum-free neuronal medium (MEM in Earle’s balanced salt containing sodium bicarbonate (0.225%), sodium pyruvate (1 mM), L-glutamine (2 mM), dextrose (0.1%), 1 X antibiotic Pen-Strep) and dissociated upon trypsin treatment. The cells were plated on poly-lysine-coated dishesat a density of 3 X 10⁶ cells/ml in non-complemented medium+serum (5%). 5’ fluoro-2’-deoxyuridine (FdU, 1mM) was added in fresh medium after 4, 7, and 10 days in culture. Astrocytes were purified by differential, vigorous shaking of the flasks 3 days after plating the dissociated brain cells. The astrocytes strongly adhere to the flask and all other cell types are removed with the medium.