EPO促进胚胎神经干细胞向神经元分化的形态学研究

目的：探讨促红细胞生成素（erythropoietin ，EPO）促进神经干细胞（neural stem cells ，NSCs）分化的形态学表现，为细胞移植治疗脑部疾病和脑损伤提供实验依据。方法取大鼠14 d胚胎脑皮质先增殖培养，后贴壁分化培养。适时镜下观察，免疫细胞化学染色，用nestin鉴定NSCs ，GFAP鉴定神经胶质细胞、MAP2鉴定神经元。选取第3代 NSCs ，向培养基中分别添加不同剂量的 EPO ，浓度分别为0．5u/ml ，5u/ml ，50u/ml ，500u/ml ，并设不添加EPO的对照组，MAP2免疫荧光共聚焦显微镜观察NSCs向神经元方向的分化。结果1）鼠胚脑皮质在无血清悬浮培养形成大量神经球，并可传代，球内细胞均表达 nestin。分化培养，可检测出MAP2或GFAP阳性细胞。2）EPO≥5u/ml各组分化培养，表达MAP2阳性细胞显著升高（P＜0．05）。结论大鼠胚胎脑皮质体外培养可获得NSCs ；适当浓度EPO可提高NSCs向神经元的分化。

Study of morphology about the promotion of erythropoietin to the differentiation of the cultured NSCs from brain cortex in vitro

Objective Methods Results Conclusion To investigate the promotion of EPO on the differenti-ation of the neural stem cells （NSCs） in vitro from cerebral cortex of pregnant SD rats .The brain cortex were iso-lated and cultured in serum-free suspension ,and then in differentiating suspension .Nestin was used to detect the NSCs .Glia fibrillary acid protein （GFAP） and microtubule-associated protein 2 （MAP2 ） were used to detect the dif-ferentiation of NSCs by immunocytochemistry .After obtained the third passage （P3 ） of NSCs ,EPO of different concentration （0.5 ,5 ,50 ,500u/ml） was added to the medium .And there was a control group without EPO .MAP2 was used to detect the neuron after the differentiation by immunocytochemistry .The results showed that the sepa-rated cells from embryonic brain cortex formed neurospheres ,in which nestin-positive cells were detected .GFAP and MAP2-positive cells were detected after differentiation culture .MAP-2-positive cells of the EPO supplementa-tion groups （＞5U/ml） showed an significant increase in the immunochemistry dying test compared with the con-trol group .The results indicate that NSCs can be obtained from the separated cerebral cortex of embryonic rats .Ap-propriate concentration of EPO can increase the rate of the neuron after the differentiation the NSCs in vitro .