NMDA对离体培养人海马神经干细胞凋亡的影响

The effect of N- Methyl -D -Aspartate on the apoptosis of human hippocampal neural stem cells in vitro

目的研究N-甲基-D-天冬氨酸（NMDA）对离体培养的人胚胎海马神经干细胞（NSCs）凋亡的影响。方法从胎龄8～12周人胚脑中分离、传代培养并鉴定海马NSCs，取传代培养的海马NSCs随机分组：①正常组；②μmol／LNMDA组；③10μmol／LNMDA组；④30μmol／LNMDA组；⑤100μmol／LNMDA组。每组设4个复孔，分别在3、7、12天取各组细胞，用Hoechst33258进行荧光染色，计算海马NSCs的凋亡率。结果在3、7、12天时，各实验组的人胚胎海马NSCs凋亡率与正常组相比均无明显差异。结论-定条件下，早期离体培养人胚胎海马NSCs对浓度为100μmol／L及其以下的NMDA有较好的耐受性。

Objective To investigate the effect of NMDA on the apoptosis of neural stem ceils （NSCs） in vitro from human embryonic hippocampus. Methods NSCs isolated from hippocampus of abortion human embryonic tissue （ embryonic age of 8 -12 weeks） were cultured and passaged in suspension. The neurospheres were identified by nestin immunocytochemistry. Passaged cells were randomized into 5 groups： normal control group, 1 μmol/L NMDA group, 10 μmol/L NMDA group, 30 μmol/L NMDA group and 100μmol/L NMDA group, with 4 wells in each group. Cells were sampled on days 3,7 and 12 for fluorescence staining with Hoechst33258 and calculation of the apoptosis rate. Results The apoptosis rate of each experimental group had no difference from that of normal control group at the appropriate time points. Conclusion Under certain conditions, NSCs cultured in vitro from human early embryonic hippocampus have good tolerance to NMDA at the concentration of 100 μmol/L and below.