Trypsin 2

Hippocampal neuronal cultures were prepared from the embryonic (E18) Sprague-Dawley rats (Charles River Laboratories) using the methods described previously (Blair et al., 2008 (link)). Briefly, female rats with 18-19 days of gestation were anesthetized with isoflurane, and embryonic pups were surgically dissected out and decapitated. Hippocampi were harvested under sterile conditions. The hippocampal tissue was enzymatically dissociated in 0.125% trypsin II (Sigma-Aldrich). Hippocampal neural cells were isolated and placed in poly-D-lysine-coated 35 mm plastic culture dishes contained 2 ml of neurobasal medium to a culture cell density of 5×10⁵ cells/ml. The cultures were maintained in neurobasal medium supplemented with B27 (2%, v/v, Invitrogen, San Diego, CA), L-glutamine (0.5mM) and 1% penicillin/streptomycin for at least 7-10 days before being used for experiments.

Hippocampal neuronal cultures were prepared from rat embryos using the methods described previously (Blair et al., 2009 (link)). Briefly, female Sprague-Dawley rats with 18–19 days of gestation were anesthetized with isoflurane, and embryonic pups were surgically dissected out and decapitated. Hippocampi were harvested under sterile conditions. The hippocampal tissue was enzymatically dissociated in 0.125% trypsin II (Sigma-Aldrich). Isolated neural cells were placed in poly-D-lysine-coated 35 mm plastic culture dishes containing 2 ml of neurobasal medium to a culture surface cell density of 5 x 10⁵/ml. The cultures were maintained in neurobasal medium supplemented with B27 (2%, v/v, Invitrogen), glutamine (0.5mM) and 1% penicillin/streptomycin for at least 7–10 days before being used for experiments.