摘要
目的:探索从胚胎小鼠额叶皮质分离培养神经干细胞的方法。方法:无菌条件下分离孕13～15d小鼠胚胎额叶皮质脑组织,经消化及机械吹打成单细胞后接种于含有B27、bFGF的DMEM/F12培养基,培养扩增;倒置显微镜观察比较生长状况;机械分离法传代;10%血清接种分化;免疫细胞化学方法染色鉴定神经干细胞及其子代细胞的分化方向,结果:培养的部分细胞体外分裂增殖,同时表达神经干细胞特异性抗原nestin,并向神经细胞和胶质细胞分化。结论:小鼠胚脑存在具有多分化潜能的神经干细胞,它们能在体外稳定培养、传代和分化,为细胞替代治疗提供了理想的细胞来源。
Objective:To explore the culture methods of neural stem cells f rom cerebral cortex of frontal lobe in embryonic mice and to Understand the NSCs biological property.Methods:cortex of frontal lobe in embryonic mice(E13-15d) were dissociated and single cell suspensions were achieved in sterile condtions.Then the cells were cultured in DMEM/F12 supplemented with bFGF a nd B27.Immunocylochemical techniques were used to odentify stem cells and their progeny;cell growth was observed by Inverted microscope;then passaged by mechanicaI methods;10% blood serum inoculated and differentiated.Result:In the serunrtree medium containing bFGFand B27,the cells from embryonic mice were divided and expanded in vitro,in which nestin antigen was expressed.In the containing serum medium,the cells were differentiated into neurons and glia.Conclusion:Neural stem cell from the cerebral cortex of frontal lobe can be cultured,passaged and differentiated in vitro.They are ideal cell sources for cell transplantation therapy.
关键词
神经干细胞
培养
分化
胎鼠
Neural stem cells
Culture
Differentiation
Embryonic mice
