不同胎龄小鼠脑组织中神经干细胞所占比例的研究

发布时间：2018-11-18 22:02

【摘要】：目的详细了解并比较不同胎龄小鼠全脑和大脑皮层组织中神经干细胞(neural stem cells,NSCs)所占比例,为后期优化分离培养NSCs提供直接量化数据。方法分离不同胎龄小鼠全脑(胎龄12.5、14、16和18d)和大脑皮层(胎龄14、16和18 d)组织,消化成单细胞悬液,贴壁培养3~4 h,细胞免疫荧光标记NSCs特异性标记物Nestin,统计分析各组中NSCs所占比例。另外,通过实时荧光定量PCR技术检测Nestin mRNA在不同胎龄(12.5、14、16和18 d)小鼠大脑皮层中的表达水平。结果细胞免疫荧光结果显示,分离培养的不同胎龄小鼠全脑和大脑皮层组织中均有Nestin阳性细胞。在分离培养的小鼠全脑组织中胎龄12.5 d组NSCs所占比例最高,为53.42%±1.57%;胎龄18 d组NSCs所占比例最低,为25.96%±1.31%,而胎龄14 d组和16 d组位于二者之间。在分离培养的不同胎龄小鼠大脑皮层组织中,胎龄14 d组NSCs所占比例最高,为33.65%±0.29%;胎龄18d组比例最低,为25.29%±0.28%,胎龄16 d组位于二者之间(26.82%±0.30%)。实时荧光定量PCR结果显示,当把胎龄12.5 d组的小鼠大脑皮层组织中Nestin mRNA表达水平设定为1,胎龄14 d组的小鼠大脑皮层组织中Nestin mRNA的相对表达量为0.83±0.04,胎龄16 d组为0.77±0.05,胎龄18 d组为0.44±0.05。因此,随着小鼠胎龄增加,小鼠大脑皮层中的Nestin mRNA的表达水平逐渐下降。结论在小鼠大脑发育过程中,随着胎龄增加,分离培养的小鼠全脑和大脑皮层组织中神经干细胞比例逐渐降低。
[Abstract]:Objective to investigate and compare the proportion of neural stem cells (neural stem cells,NSCs) in the whole brain and cerebral cortex of mice of different gestational ages in order to provide direct quantitative data for optimizing isolation and culture of NSCs in later stage. Methods the whole brain and cerebral cortex of mice of different gestational ages were isolated and digested into single cell suspension for 3 hours. The specific NSCs marker Nestin, was labeled with cellular immunofluorescence. The proportion of NSCs in each group was analyzed statistically. In addition, real-time fluorescence quantitative PCR was used to detect the expression of Nestin mRNA in the cerebral cortex of mice at different gestational ages (12.5U 1416d and 18d). Results the results of cellular immunofluorescence showed that there were Nestin positive cells in the whole brain and cerebral cortex of mice of different gestational ages. In the whole brain of the isolated and cultured mice, the proportion of NSCs was the highest (53.42% 卤1.57) in the 12.5-day gestational age group. The proportion of NSCs in 18 d gestational age group was the lowest (25.96% 卤1.31), while the gestational age 14 d group and 16 d group were between them. The proportion of NSCs in the 14 d gestational age group was the highest (33.65% 卤0.29) in the cerebral cortex of mice of different gestational ages. The lowest proportion was 25.29% 卤0.28 in 18 d gestational age group, and 26.82% 卤0.30% in 16 d gestational age group. The results of real-time fluorescence quantitative PCR showed that the relative expression of Nestin mRNA was 0.83 卤0.04 in the cerebral cortex of mice at gestational age of 14 days, when the expression level of Nestin mRNA was set to 1 in the cerebral cortex of the 125th day gestational age group, and the relative expression level of Nestin mRNA in the cerebral cortex was 0.83 卤0.04 in the 14-day gestational age group. The gestational age was 0.77 卤0.05 in 16 d group and 0.44 卤0.05 in 18 d gestational age group. Therefore, with the increase of gestational age, the expression of Nestin mRNA in cerebral cortex of mice decreased gradually. Conclusion during the development of mouse brain, the proportion of neural stem cells in the whole brain and cerebral cortex of the isolated and cultured mice gradually decreased with the increase of gestational age.
【作者单位】：昆明医科大学分子临床医学研究院云南省干细胞和再生医学重点实验室;深圳市宝安区人民医院;南方医科大学珠江医院神经外科广东神经外科研究所广东省脑功能修复与再生重点实验室;昆明医科大学基础医学院;澳大利亚莫纳什大学免疫与干细胞研究中心;
【基金】：云南省高端人才引进计划(编号:20080A004) 云南省干细胞和再生医学重点实验室(编号:2015DG027)
【分类号】：R-33

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