Primary rat cortical neuron cultures

Primary rat cortical neuron cultures were obtained as previously published by our group [18]. In short, cortical neurons were obtained from brain rats at embryonic days 18-19 (E18-19) and cultured in high glucose (4.5 g.L^-1) Neurobasal medium with 500 μM L-Glutamine, 2 % B27 supplement, 100 U.mL^-1 penicillin and, 100 μg.mL^-1 streptomycin (all from Gibco), with no growth factors, on poly-D-Lysine coated cell culture plates. Cortical neurons were maintained at 37° C in a humidified incubator with 5 % CO₂/air for 8 days and the medium was replaced every fourth day by aspirating half of the medium from each well and replacing it with fresh medium.