目的探讨新生小鼠端脑神经干细胞诱导分化为运动神经元的可能性,并探索新的运动神经元诱导因子。方法用悬浮培养法从新生小鼠端脑分离培养神经干细胞,按诱导因素的不同分为3组:组1为对照组,诱导因素为生长培养基+5%胎牛血清(FBS);组2为诱导因子组,诱导因素为生长培养基+5%FBS+视黄酸(RA)+Shh+联丁酰基环磷酸腺苷(dbcAMP);组3为骨骼肌细胞培养液组,诱导因素为骨骼肌细胞生长过的培养液。用双重免疫荧光方法检测分化细胞的微管相关蛋白2(MAP2)和同源蛋白(HB9)的表达,以验证运动神经元的分化,每组随机选取12个标本计数。结果分化培养中检测到MAP2和HB9共表达的运动神经元,组1的运动神经元分化比例为1%;组2的分化比例为4.7%;组3的分化比例为2.9%。与组1相比有显著的统计学差异。结论新生小鼠端脑神经干细胞能诱导分化为运动神经元;骨骼肌细胞可能分泌运动神经元诱导因子。 Objective To investigate the possibility of neural stem cells differentiating into motor neurons in newborn mice and to explore new motor neuron - induced factors. Methods The neural stem cells were isolated from neonatal mice by suspension culture and divided into 3 groups according to the inducing factors: group 1 was control group, the inducer was growth medium + 5% fetal bovine serum (FBS); group 2 As induction factor group, the induction factors were growth medium + 5% FBS + RA + Shh + bcAMP; group 3 was a culture medium of skeletal muscle cells, and the inducing factors were skeletal muscle cells Growth medium. Differentiation of microtubule-associated protein 2 (MAP2) and homologous protein (HB9) in differentiated cells was detected by double immunofluorescence to verify the differentiation of motor neurons. Twelve specimens were randomly selected from each group. Results Motor neurons co-expressed by MAP2 and HB9 were detected in differentiation culture. The ratio of motor neuron differentiation in group 1 was 1%. The differentiation ratio in group 2 was 4.7%. The differentiation ratio in group 3 was 2.9%. There was a significant statistical difference compared to group 1. Conclusion Neonatal mouse embryonic neural stem cells can differentiate into motor neurons; skeletal muscle cells may secrete motoneurons.