摘要
目的:建立胎鼠大脑皮层神经干细胞体外分离、培养、分化及鉴定的方法,探讨神经干细胞的基本生物学特性。方法:取孕14 d胎鼠大脑皮层,吸管吹打机械分离制成单细胞悬液,采用无血清培养和胎牛血清诱导分化,应用免疫荧光技术鉴定神经干细胞及其分化的子代细胞。结果:从胎鼠大脑皮层分离的细胞呈神经巢蛋白(Nestin)免疫阳性并能在体外传代培养,血清诱导分化后的细胞可表达神经元、星形胶质细胞的特异性抗原微管相关蛋白2和胶质纤维酸性蛋白,诱导分化成的神经元能表达突触的特异性标志Synatophymin。结论:分离和培养的细胞能表达神经干细胞的特异性标志物Nestin,并且具有很强的增殖能力和多向分化潜能,属于中枢神经系统的神经干细胞。
Objective： To establish a method for isolation, cultivation, differentiation induction, and identification of neural stem cells （NSCs） from embryonic rat in vitro, and explore the basic biological features of these cells. Methods： The mesencephalon of rat with conceptus age of 13 days was taken out and dissociated into single cell suspension mechanically, and then cultured in serum-free medium. Fetal bovine serum was used to induce cell differentiation. The isolated cells and their progeny were observed by using immunofluorescence technique. Result： The isolated cells showed nestin positive reaction and could be cultured in vitro in series. After being induced by fetal bovine serum, microtubuleassociated protein 2 （MAP-2） and glial fibrillary acidic protein （GFAP） which are specific antigens in neuron and astrocyte expressed in these cells, and synatophymin, a specific signal of synapse, was also expressed in the differentiated neurons. Conclusion： The isolated and cultured cells are NSCs of central nervous system, and possess strong capability of proliferation and potency of multi-directional differentiation.
出处
《贵阳医学院学报》
CAS
2008年第4期347-350,共4页
Journal of Guiyang Medical College
基金
贵州省科技厅基金项目(20043002)
贵阳市科技局项目(2004第26号)
