Primary Cultures of Rat Cortical Cells
Primary cultures of cortical cells containing neuronal and non-neuronal cells were prepared from the brains of SD rat embryos at 17 d gestation as described previously.22–24) In brief, cerebral cortices of the embryos were dissected and mechanically dissociated into single cells by trituration using fire-polished Pasteur pipettes. The isolated cells were seeded in MEM supplemented with 5% FBS, 5% HS and 1% antibiotic-antimycotic agent at a density of 1–6×105 cells/well of a 24- or 96-well culture plate (BD Falcon, Franklin Lakes, NJ, U.S.A.) or at a density of 6×106 cells per a 35 mm culture dish (Corning, NY, U.S.A.) pre-coated with laminin and poly-L-lysine. The cells were maintained in an incubator at 37°C with a humidified atmosphere of 95% air/5% CO2. Proliferation of non-neuronal cells in the culture was arrested by the treatment with 10 µM cytosine arabinoside at 7 d after seeding of cells. All experiments were carried out at 10–11 d after plating.
Primary Cultures of Rat Cortical CellsPrimary cultures of cortical cells containing neuronal and non-neuronal cells were prepared from the brains of SD rat embryos at 17 d gestation as described previously.22–24) In brief, cerebral cortices of the embryos were dissected and mechanically dissociated into single cells by trituration using fire-polished Pasteur pipettes. The isolated cells were seeded in MEM supplemented with 5% FBS, 5% HS and 1% antibiotic-antimycotic agent at a density of 1–6×105 cells/well of a 24- or 96-well culture plate (BD Falcon, Franklin Lakes, NJ, U.S.A.) or at a density of 6×106 cells per a 35 mm culture dish (Corning, NY, U.S.A.) pre-coated with laminin and poly-L-lysine. The cells were maintained in an incubator at 37°C with a humidified atmosphere of 95% air/5% CO2. Proliferation of non-neuronal cells in the culture was arrested by the treatment with 10 µM cytosine arabinoside at 7 d after seeding of cells. All experiments were carried out at 10–11 d after plating.
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