小鼠胚胎皮质、海马来源的神经干细胞在体外增殖特性的比较研究

The differences of neural stem cells proliferation between cortex and hippocampus in embryonic mice

目的探讨皮质、海马来源的神经干细胞增殖特性的异同,为神经干细胞基础和临床研究在取材部位上提供参考资料。方法无菌条件下分别分离E11-15天小鼠胚脑皮质和海马,经胰酶消化及机械吹打制成单细胞悬液后,在含B27和bFGF的DMEM/F1224孔板中培养扩增,倒置显微镜观察比较生长状况,免疫细胞化学染色技术鉴定神经干细胞,采用BrdU掺入法检测不同部位来源NSCs的增殖情况。结果小鼠胚胎脑皮质与海马均存在神经干细胞,皮质部位神经干细胞比海马部位神经干细胞更易培养,也更易成球,前者神经球数目、BrdU阳性细胞率也明显高于后者。结论同样条件下皮质神经干细胞比海马神经干细胞更易培养和增殖。

Objective To explore the differences of cultivation and proliferation between cortical neural stem cells （C-NSCs） and hippocampal neural stem cells（H-NSCs）. Methods The cortex and hippocampus of 11-15 days mice embryo were dissociated, and single cell suspensions were achieved by trypsin digestion and mechanical dissociation in sterile condition. These cells were planted into DMEM/F12 culture medium containing B27 and bFGF. Reverse phase-contrast microscopy and inverted microscope were applied to observe and compare the differences of the two kinds of NSCs. Immunocytochemical technique was applied to detect the expression of nestin,proliferative ability of NSCs was detected with 5-bromo-2＇deoxyuridine （BrdU）. Results The positive NSCs were found to be existed both in cortex and hippocampus of embryonic mice, with the easier plantation for C-NSCs than for H-NSCs. The density of NSCs and the rate of BrdU-positive cells were much higher in cortex than in hippocampus. Conclusion The proliferation activity of C-NSCs is higher than that of H-NSCs at the same condition.