Primary cultures of hippocampal CGP41251 were prepared according to established procedures with slight modifications [15]. Briefly, hippocampal tissue was obtained from E18 mouse embryos, dissected, and dissociated with the Papain Dissociation System (PDS; Worthington Biochemical Corporation, Lakewood, NJ, USA). Neurons were maintained in Neurobasal medium (Gibco, Grand Island, NY, USA) supplemented with 2% B-27, 1% N2, 0.5 mM glutamine, 100 U/ml penicillin, and 100 mg/mL streptomycin, and grown on poly-l-lysine (100 mg/mL) coated plastic dishes. The cells were incubated in 95% air/5% CO2 in a humidified incubator at 37 °C.
Human neuroblastoma SH-SY5Y cells [16] were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum, 2 mM glutamine, 100 U/ml penicillin, 100 U/ml streptomycin in a humified atmosphere of 5% CO2/95% air at 37 °C. For differentiation, proliferating SH-SY5Y cells were plated and then cultured in Neurobasal-B27 medium (Gibco, Grand Island, NY) supplemented with 2 mM dibutyril cyclic AMP and 1 mM glutamine for 7 days.