Primary cortical neurons are isolated from human fetal brain tissue (elective abortus specimens) routinely obtained from our neural tissue core facility and are cultured as described [38]. The tissues are obtained in full compliance with the ethical guidelines of both the National Institutes of Health and the University of Nebraska. Briefly, we incubated dissociated tissue with 0.25% trypsin for 30 min, neutralized with 10% fetal bovine serum, and further dissociated by triturating. Resulting single-cell suspension was cultured on poly-D-lysine-coated plates in neurobasal media containing 0.5 mM glutamine, 50 μg/ml penicillin and streptomycin, and supplemental B27. Neuronal purity (>75%) was assessed by antibodies (Abs) to neuronal markers, microtubule-associated protein 2 (MAP-2, Boehringer, 1:50) and neurofilament (NF: Dako, 1:100), and astrocytic marker, glial fibrillary acidic protein, GFAP (Dako, 1:100).
