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Briefly, embryos were surgically removed and the cortical areas were dissected from the cerebral tissue in Hanks balanced salt solution (HBSS, Gibco, Life Technologies, cat.# 14170088), freed of meninges, digested with 0.25% trypsin (Gibco, Life Technologies, cat.# 15090-046) for 15 min at 37C, dissociated by trituration and plated (10 6 cells/dish) on 35 mm poly-L-lysine-coated wells (Poly-Lysine hydrobromide, Sigma-Aldrich, cat.# P5899) in Earles minimum essential medium (MEM, Gibco, Life Technologies, ca.#t 41090-028) containing 10% foetal bovine serum (Gibco, Life Technologies, cat.# 16000-044) and 2% glucose (Sigma-Aldrich, cat.# G7021)