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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 similarities and differences of proliferation characteristics of neural stem cells derived from cortex and hippocampus and to provide reference materials for the basic and clinical research of neural stem cells in the derived site. Methods The embryonic cortex and hippocampus of E11-15 mice were isolated under aseptic conditions. After single cell suspension was prepared by trypsin digestion and mechanical blowing, the cells were cultured and expanded in DMEM / F1224 plates containing B27 and bFGF. Inverted microscope was used to compare the growth status. Immunocytochemical staining was used to identify neural stem cells. BrdU incorporation was used to detect the proliferation of NSCs from different sites. Results Neural stem cells existed in the cerebral cortex and hippocampus of mouse embryos. Neural stem cells of cortical neurons were more easily cultured than neural stem cells in hippocampus and more easily formed into spheres. The number of neurospheres and BrdU positive cells in the former were also significantly higher than those in the latter. Conclusion Under the same conditions, cortical neural stem cells are more easily cultured and proliferated than hippocampus neural stem cells.