新生小鼠端脑组织神经干细胞诱导分化为胆碱能神经元的研究

目的探讨新生小鼠端脑组织神经干细胞是否能够分化成胆碱能神经元。方法取新生小鼠端脑组织．用无血清方法分离培养神经干细胞；用克隆培养的方法检验培养细胞的干细胞特性；用免疫荧光细胞化学的方法检测神经干细胞标志巢蛋白（nestin）及干细胞诱导分化后神经元标志微管相关蛋白2（MAP2）、星形胶质细胞标志胶质纤维酸性蛋白（GFAP）、胆碱能标志胆碱乙酰转移酶（CHAT）；比较不同的诱导分化条件（5％胎牛血清、5％胎牛血清＋碱性成纤维细胞生长因子）对胆碱能神经元分化的影响。结果从新生小鼠端脑组织分离培养出具有自我更新、扩增能力的神经球；各培养基中神经球均为nestin阳性。诱导分化后均能够产生MAP2阳性神经元、GFAP阳性星形胶质细胞以及ChAT阳性的胆碱能神经元。分化培养中加入碱性成纤维细胞生长因子能够提高胆碱能神经元分化的比例。结论新生小鼠端脑组织神经干细胞能够分化成胆碱能神经元。

Differentiation of neonatal mouse telencephalon-derived neural stem cells into cholinergic neurons

Objective To explore whether neonatal mouse telencephalon-derived neural stem cells could be induced to differentiate into cholinergic neurons. Methods Neural stem cells were isolated from neonatal mouse telencephalon and cultured in serum-free medium. Clonal culture was carried out to demonstrate that the isolated cells met the standard of stem cells. Fluorescence immunocytochemistry was used to examine the expression of neural stem cell marker nestin, neuronal marker MAP2, astrocyte marker GFAP and cholinergic marker CHAT. The effect of different inductive conditions, such as 5% fetal bovine serum, 5% fetal bovine serum together with basic fihrohlast growth factor, on the differentiation into cholinergic neurons was compared. Results Self-renewal and expandable neural stem cells were obtained from neonatal mouse telencephalon and grown as floating neurospheres. Neurospheres in all cultures showed nestin positive and could yield GFAP-positive astrocytes and MAP2-positive neurons including CHAT-positive cholinergic neurons in differentiation cultures. The addition of basic fihrohlast growth factor in differentiation culture could increase the proportion of cholinergic neurons. Conclusion Neonatal mouse telencephalon-derived neural stem cells can be induced to differentiate into cholinergic neurons.