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目的研究N-甲基-D-天门冬氨酸(NMDA)对离体培养的人胚胎海马神经干细胞(NSCs)增殖的影响。方法取胎龄8～12周人胚脑海马,分离、培养、传代、鉴定NSCs,取传代培养的NSCs随机分为以下5组:对照组、1μmol/L NMDA组、10μmol/L NMDA组、30μmol/L NMDA组、100μmol/L NMDA组。每组取8孔,对照组使用正常培养基培养,各NMDA组加入不同剂量的NMDA,放在培养箱中继续培养。各组细胞分别在3 d、7 d和12 d后采用MTT法测定吸光度(A)值。结果在NMDA作用3 d和7 d时,100μmol/L NMDA组的A值显著高于对照组(均P<0.05);其他各组与对照组相比,以及不同浓度NMDA组间相比,差异均无统计学意义(均P>0.05)。NMDA作用12 d时,30μmol/L和100μmol/L NMDA组的A值显著高于对照组(均P<0.05);其他各组与对照组相比以及不同浓度NMDA组间相比,差异均无统计学意义(均P>0.05)。结论一定条件下NMDA可以促进离体培养人胚胎海马NSCs增殖。
Objective To investigate the effect of N-methyl-D-aspartate (NMDA) on the proliferation of cultured human embryonic hippocampal neural stem cells (NSCs). Methods The hippocampus of human embryonic brain at 8-12 weeks gestation was isolated, cultured, passaged and identified. NSCs cultured in subculture were randomly divided into the following 5 groups: control group, 1μmol / L NMDA group, 10μmol / L NMDA group, 30μmol / L NMDA group, 100μmol / L NMDA group. Each group take 8 holes, the control group using normal culture medium, each NMDA group added different doses of NMDA, placed in the incubator and cultured. The absorbance (A) value of each group of cells was measured by MTT method after 3 days, 7 days and 12 days respectively. Results The A value of NMDA group was significantly higher than that of control group at 3 d and 7 d of NMDA treatment (all P <0.05). Compared with control group and NMDA group, No statistical significance (all P> 0.05). At 12 days after NMDA administration, the values of A in 30μmol / L and 100μmol / L NMDA group were significantly higher than those in control group (all P <0.05). There was no significant difference in other groups compared with control group and NMDA group Statistical significance (all P> 0.05). Conclusion Under certain conditions, NMDA can promote the proliferation of human embryonic hippocampal NSCs cultured in vitro.