Hippocampi were dissected and dissociated in papain. Cells were plated at a density of 225,000 viable cells/well in 24-well glass-bottom plates (Mattek, #1.5 glass coverslips), pre-coated with Matrigel (BD Biosciences). Cells were cultured in growth medium (28 mM glucose, 2.4 mM sodium bicarbonate, 100 μg/mL transferrin, B-27 supplement (1X, Invitrogen), 500 μM L-glutamine, 50 units/mL penicillin, 50 mg/mL streptomycin, 5% fetal bovine serum in MEM).
