NGF诱导大鼠胚脑皮层和隔区神经干细胞向神经元和AChE阳性神经元分化的作用

目的:探讨大鼠胚脑皮层和隔区神经干细胞在NGF的诱导下分化为神经元和AChE阳性神经元的情况。方法:应用无血清培养技术,分别从SD大鼠胚脑皮层和隔区分离克隆神经干细胞,然后在含或不含NGF的培养液中分化14d。用MAP-2免疫荧光和AChE组化技术检测神经干细胞分化为神经元和AChE阳性神经元的情况。并对MAP-2阳性神经元的分化率、AChE阳性神经元数及其它们的细胞面积和周长进行图像处理;用STATA7.0统计软件进行方差分析和两两比较。结果:隔区和皮层NGF组MAP-2阳性神经元分化率高于隔区和皮层对照组;其细胞面积除隔区NGF组与皮层NGF组之外,其它各组间差异均有统计学意义;而其周长各组之间差异均有统计学意义。AChE阳性神经元的数量及细胞面积和周长,隔区NGF组最佳,皮层NGF组次之,隔区对照组再次之,皮层对照组较差。三项指标各组间差异有统计学意义。结论:NGF可诱导大鼠胚脑皮层和隔区神经干细胞向神经元分化;隔区神经干细胞较皮层神经干细胞更易于向AChE阳性神经元分化;这种区域特异性可在NGF诱导下发生改变。

Effects of NGF on inducing the rat embryonic neural stem cells derived from cortex and septal region to differentiate into neurons and AChE positive neurons in vitro

Objective: To study the effects of NGF on inducing the rat embryonic neural stem cells (NSCs) derived from cortex and septal region differentiating into neurons and AChE positive neurons in vitro. Methods: The NSCs were obtained from the rat fetal cortex and the septal region by the technique of serum-free culture respectively. The NSCs were differentiated in the medium with or without NGF for 14 days. The MAP-2 and the AChE positive neurons were detected by using immunofluorescence and histochemistry respectively. The image processing about the differentiation rates of MAP-2 positive neurons, the number of AChE positive neurons, the dimensions and perimeters of the neurons were employed. Then, the analysis of variance was made and Newman-Keuls test was used by STATA 7.0 statistical software. Results: The differentiation rates of the MAP-2 positive neurons in septal region NGF group and the cortex NGF group were superior to those in septal control and the cortex region groups. As far as dimensions of the MAP-2 positive neurons were concerned, there were significant differences among other groups except for between the septal region NGF group and the cortex NGF group. For the perimeters of MAP-2 positive neurons, there were significant differences among all groups. For the results about the number, dimension and perimeter of AChE positive neurons, septal region NGF group was the best, cortex NGF group the second, septal region control group the third and cortex control group was the worst. There were significant differences among all groups for above 3 indexes. Conclusions: NGF has the effects on inducing neural stem cells derived from rat fetal cortex and septal region to differentiate into neurons. The neural stem cells derived from septal region is more liable differentiate to AChE positive neurons than those which come from cortex; This regional specificity about NSCs differentiating into AChE positive neurons can be exchanged by the inducing of NGF.