海马神经元培养.docVIP

海马神经元培养 (1)材料 ①孕鼠:海马:孕18d,皮层或纹状体:孕16d ②Neurobasal medium (Gibco-BRL, cat. no. 21103) L-Glutamine (Gibco-BRL, cat. no. 25030) 谷氨酰胺 Glutamic acid (Sigma, cat. no. G-1626) 谷氨酸 B27 (Gibco-BRL, cat. no. 17504) ③Poly-D-lysine (mol wt 30,000—70,000) (Sigma, cat. no. P-7280)多聚赖氨酸 ④Hank’s balanced salt solution (HBSS) (Gibco-BRL, cat. no. 14025) ⑤HBSS without Ca2+, Mg2+ (Gibco-BRL, cat. no. 14175) ⑥trypsin 胰酶 ⑦NaHCO 3 ⑧Na pyruvate (Gibco-BRL, cat. no. 11840)丙酮酸钠 ⑨Trypan blue, 0.4% 台盘蓝 ⑩巴氏吸管,前端用火抛光 刻度吸管 离心管 闪烁瓶 玻璃培养皿:小:3套 大:2套 冰袋、纱布 手术器械、筛网 培养瓶或培养板 (2)步骤 ①准备 a.配制试剂 i) Poly-D-l