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目的 观察无血清培养法诱导小鼠胚胎干细胞(Embryome stem cell,ESCs)ESCs体外分化为神经干细胞(Neural stem cell, NSCs)的诱导效率。方法 采用无血清培养法诱导小鼠ESCs 体外分化为NSCs,对诱导不同时间点的细胞行免疫荧光,RT PCR检测nestin的表达。结果 诱导48 h开始出现nestin mRNA的表达,诱导72 h出现nestin 蛋白表达。诱导120 h nestin 的表达达高峰,阳性细胞率为(70.3±8.02)%。Nestin阳性细胞呈球形生长,可不断增殖。结论 本诱导方法可获得较高纯度的NSCs,所获得的神经干细胞与脑内来源的NSCs有相似的生物学特性。
Objective To observe the induction efficiency of ESCs differentiated into neural stem cells (NSCs) from mouse embryonic stem cells (ESCs) by serum-free culture. Methods Mouse ESCs were induced to differentiate into NSCs by serum-free culture in vitro. Immunofluorescence was performed on cells induced at different time points. The expression of nestin was detected by RT-PCR. Results The expression of nestin mRNA began to appear 48 h after induction, and the expression of nestin protein appeared 72 h after induction. The expression of nestin peaked at 120 h, the positive rate was (70.3 ± 8.02)%. Nestin-positive cells are spherical growth, can continue to proliferate. Conclusion The method of this induction method can obtain higher purity of NSCs. The obtained neural stem cells have similar biological characteristics to NSCs derived from brain.