摘要
目的:探索体外培养大鼠神经干细胞(Neural stem cells,NSCs)的方法和实验条件,建立一种较为可靠的神经干细胞培养方法。方法:分别选取孕14～16dSD胎鼠和新生1d的SD大鼠进行神经干细胞培养,采用单纯机械吹打和机械吹打加酶消化两种方法进行消化,对培养出的细胞进行神经干细胞、神经元、神经胶质细胞的免疫细胞化学和免疫荧光鉴定。结果:用2种来源的大鼠培养出的原代细胞,均表达Nestin,诱导分化后的细胞表达GFAP和MAP-2。结论:大鼠神经干细胞的原代分离与培养的关键是单细胞悬液的获得,消化液浓度太高或消化时间太长,容易因消化过度致细胞损伤甚至死亡。胎鼠来源者神经球形成早,数量多,增殖活力更强,但原代取材过程易被污染。新生鼠来源者鼠源易获得,组织取材过程操作简单,但神经干细胞成球率低。
Objective:To establish the method of culturing neural stem cells(NSCs) from rat brain. Methods:E14～16 SD embryonic rat brain and postnatal one-day(P1) rat brain were respectively chose to culture neural stem cells,and two digestive methods that simple mechanical digestion and mechanical combined with chemical digestion were used to determine the best conditions in primary culture.Immunocytochemistry and immunofluorescence were used to identify NSCs,neurons and neuroglial cells. Results:Both the primary cells from two different kinds of rat brains expressed nestin antigen and the differentiated cells expressed GFAP and MAP-2 antigen.Conclusion:The key point of primary NSCs culture was obtaining the monoplast suspension.Overdigestion led to cell injury even death. NSCs from embryonic rat brain had better proliferation ability and could form cell clones earlier and more than NSCs from newborn rat brain,but they were easier to be contaminated. Obtaining newborn rat brain and isolation NSCs from newborn rat brain were easier,but NSCs from newborn rat brain had worse proliferation ability.
出处
《重庆医科大学学报》
CAS
CSCD
2008年第z1期44-47,共4页
Journal of Chongqing Medical University
关键词
神经干细胞
细胞培养
免疫荧光
免疫生化
Neural stem cells
Cell culture
Immunofluorescence
Immunocyto chemistry
