小胶质细胞的培养及鉴定

目的:获得高纯度培养原代小胶质细胞的方法并检测Notch信号通路相关分子在小胶质细胞的表达情况。方法:取胎鼠利用反复机械振摇纯化分离小胶质细胞;利用流式细胞仪,根据CD11b及MHCII的表达水平对分离的小胶质细胞纯度进行鉴定;利用qPCR及琼脂糖凝胶电泳检测小胶质细胞中Notch通路相关分子的表达情况。结果:利用5只胎鼠采取反复机械振摇的方法可较稳定的获得1.1×106个的小胶质细胞,流式细胞术结果显示细胞纯度高达97.77%,并在小胶质细胞中检测到Notch相关分子的表达。结论:利用胎鼠反复机械振摇法可以获得较高纯度及产量的小胶质细胞,小胶质细胞表达Notch信号通路。

Cultivation and Identification of PrimaryMicroglia

Objective: To find a method to get the primary microglia with high purity and examine the expression of Notchsignal-associated molecules in microglia. Methods: Fetal mice cerebral cortex were used to isolate microglia by repetitively mechanicalshaking. The purity of isolated cells was identified by expression of CD11b and MHCII using flow cytometry. The expression of Notchsignal-associated molecules was examined by qPCR and gel electrophoresis. Results: 1.1×106 microglial cells with high purity (97.77%)could be steadily produced from 5 fetal mice by repetitively mechanical shaking and the Notch signal-associated molecules wereexamined in microglia. Conclusions: Primary microglia with high production and purity can be obtained by repetitively mechanicalshaking from fetal mice cerebral cortex, and the Notch signaling pathway is expressed in microglia.