促红细胞生成素体外培养鼠胚脑皮质神经干细胞的分化

背景:促红细胞生成素最早作为生长因子被认识,近些年来其对中枢神经系统保护作用的体内研究较多。目的:观察促红细胞生成素对体外培养大鼠胚脑皮质神经干细胞凋亡及分化的影响。方法:无菌条件下取孕14dSD大鼠胚脑皮质,体外先悬浮增殖培养后贴壁诱导分化。巢蛋白免疫细胞荧光染色检测神经干细胞,以微管相关蛋白2、神经胶质原纤维酸性蛋白检测神经干细胞分化。取第3代神经干细胞向培养基中添加0.5,5,50,500U/mL的促红细胞生成素,另设不添加促红细胞生成素的对照组。应用半胱氨酸天冬氨酸蛋白酶3检测神经干细胞凋亡,微管相关蛋白2检测神经干细胞向神经元方向的分化。结果与结论:加入≥5U/mL促红细胞生成素,神经球内半胱氨酸天冬氨酸蛋白酶3表达显著下降(P<0.01),分化培养后微管相关蛋白2阳性细胞较对照组明显升高(P<0.01)。提示促红细胞生成素可降低体外培养鼠胚脑皮质神经干细胞的凋亡率,促进其神经干细胞向神经元方向分化。

Differentiation of rat embryonic cerebral cortex neural stem cells cultured by erythropoietin in vitro

BACKGROUND:Erythropoietin(FPO) is early known as a kind of growth factor.In recent years,there are many studies about the protective effect of erythropoietin on the central nervous system in vivo.OBJECTIVE:To explore the effects of EPO on the apoptosis and differentiation of rat embryonic cerebral cortex neural stem cells in vitro.METHODS:The embryonic cerebral cortex were isolate from Sprague-Dawley rats that pregnant for 14 days under sterile conditions,the cells were cultured and differentiated in the suspension medium and then adherent induction.The neural stem cells were detected by nestin immune cell fluorescence staining,microtubule-associated protein 2(MAP-2) and glia fibrillary acid protein was used to detect the differentiation of neural stem cells.The third passage of neural stem cells were obtained and 0.5,5,50,500 U/mL EPO were added into the medium,and in the control group,there was no EPO adding.Caspase-3 was used to detect the apoptosis of neural stem cells,and MAP-2 was used to detect the differentiation of neural stem cells to neuron.RESULTS AND CONCLUSION:The expression of the caspase-3 in neural sphere was decreased after added EPO(≥5 U/mL),and the expression of the MAP-2 positive cells was increased obviously after differentiation(P < 0.01).The present results suggest that EPO can decrease the apoptosis of the neural stem cells and promote neural stem cells to differentiate into neurons in vitro.