新生小鼠端脑神经干细胞可诱导分化为运动神经元

Motor neuron differentiation from neonatal mouse telencephalon neural stem cells

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 explore the possibility of motor neural stem cells and the novel inducing factors. Methods neuron differentiation from neonatal mouse telencephalon Neural stem cells were isolated from neonatal mouse telencephalon using floating culture methods. Three differentiation groups were divided according to different inducing factors. Group 1 containing growth medium ＋ 5% fetal bovine serum （FBS） served as control. Group 2 containing growth medium ＋5% FBS ＋ retinoid acid （RA） ＋ Shh ＋ dbcAMP was inducing factor group. Medium collected from skeletal muscle cultures was used for motor neuron induction in Group 3. Double immunofluoesence staining of microtubule associated protein 2 （MAP2） and borneo box（HB9） were used to detect the differentiation of motor neuron. Twelve samples were selected randomly for cell counting. Results Motor neurons with MAP2 and HB9 coexpression were found in differentiation cultures. The ratio of motor neuron differentiation was 1% in Group 1, 4.7% in Group 2 and 2.9% in Group 3. The difference between Group 2 or Group 3 and Group 1 was of statistic significance. Conclusion Neonatal mouse telencephalon neural stem cells can differentiate into motor neurons. Skeletal muscle cells might secret motor neuron inducing factors.