比较四种消化方法对神经干细胞传代增殖及活性的影响

Compare the effect of four kinds of digestion methods to the proliferation and activity of the neural stem cell passage

目的:观察4种消化方法对体外培养的神经干细胞(N SC s)增殖及活性的影响。方法:原代培养新生大鼠海马N SC s,采用机械消化、0.25%单纯胰酶、0.25%胰酶+0.02%EDTA和0.02%EDTA 4种不同消化方法对N SC s球进行传代培养,经四唑盐比色法(M TT法)法检测不同方法对N SC s活性及增殖能力的变化。结果:与机械消化相比,0.25%单纯胰酶组,0.25%胰酶+EDTA组消化后的大鼠海马N SC s球长时间不聚集,细胞增殖停顿,而单纯EDTA组则明显提高了N SC s的增殖及活性,其余各组差异无统计学意义。结论:随培养时间延长,单纯0.02%EDTA组能够显著提高体外培养的大鼠海马N SC s的增殖水平及活力,且对细胞的后续损伤相对较小。

Objective：To observe the four kinds of digestion of NSCs in vitro proliferation and activity.Methods：Primary culture of neonatal Wistar rat hippocampal NSCs,mechanical separation,0.25% pure trypsin,0.02% EDTA and 0.25% trypsin ＋0.02% EDTA4 different digestion methods NSCs ball subculture,the four methyl thiazolyl tetrazolium assay（MTT） method for detection of four kinds of digestive activity and proliferation of NSCs changes.Results：Compared with the mechanical digestion,0.25% trypsin alone group,0.25% trypsin ＋ EDTA group after digestion in rat hippocampal NSCs do not gather the ball for a long time,cell proliferation stopped,and proliferation of NSCs and activity were significantly increased by pure EDTA group,which makes no significant difference in the other groups.Conclusion：With the cultivate time extended,pure 0.02% EDTA group can significantly improve the in vitro proliferation of hippocampal NSCs level and vitality,and the subsequent damage to the cells is relatively small.