神经干细胞相关抗原在SD大鼠脑及O-2A谱系细胞的表达研究

发布时间：2017-12-31 08:02

本文关键词：神经干细胞相关抗原在SD大鼠脑及O-2A谱系细胞的表达研究　出处：《苏州大学》2012年硕士论文　论文类型：学位论文

【摘要】：第一部分神经干细胞相关抗原在SD大鼠脑组织的表达目的观察分析NG2、CD15与nestin在不同时期SD大鼠脑组织中的表达变化情况。方法将出生后7天和3个月的两组SD大鼠麻醉后进行灌注、固定、取脑，4%多聚甲醛后固定4小时后入10%、20%蔗糖脱水后，依照徐州医学院王平宇编著的《大鼠脑读片提要及图谱》进行冰冻切片。对神经干细胞相关抗原（nestin，CD15，NG2）和O-2A谱系特异性标记物A2B5分别进行免疫荧光组织化学染色研究，观察表达差异。结果7天组和3个月组的脑组织中，脑皮质和脑髓质中几乎观察不到nestin和CD15阳性细胞，而在SVZ和海马齿状回附近集中表达，表达量随着鼠龄的增长略微有些减少；NG2在不同脑区均有发现，与早期脑组织不同的是，成熟的脑组织在室管膜下区NG2表达减少，而在SVZ区周围的实质部分大量表达NG2。结论神经干细胞发育过程中会向脑组织周边迁移，表面抗原nestin消失的最快，其次是CD15，NG2发育过程中消失的最慢或者不消失。第二部分神经干细胞相关抗原在O-2A谱系胶质细胞中的表达目的观察分析神经干细胞相关抗原NG2、CD15与nestin在O-2A谱系胶质细胞中的表达变化情况。方法根据O-2A谱系细胞的生长时间差异及细胞的粘附特性不同，采用振荡法和差速贴壁法，并结合使用含生长因子的无血清培养基以获得纯化的O-2A祖细胞，将O-2A祖细胞用生长因子培养基诱导分化为T2A后，对培养1d和2d的O-2A和培养纯化1d和3d的T2A分别进行nestin，CD15，NG2和A2B5的免疫荧光检测。结果nestin在OPC（1d,2d）表达较强，在T2A培养早期（1d）也有表达但已稍减少（仅有70-80%表达），，在T2A分化成熟阶段（3d后）几乎不见nestin表达。同样，CD15在OPC比T2A表达强，但有意思的是，在分化成熟的T2A中CD15虽表达减少但并未完全不表达。NG2在O-2A谱系（OPC、T2A）中均有较强表达。结论O-2A谱系胶质细胞在中枢神经系统发育过程中，其神经干细胞标记物的表达，随着细胞由未成熟至成熟状态而逐渐减弱，但不同神经干细胞标记物之间表达规律有差异。
[Abstract]:The first part of the expression of neural stem cell related antigen in the brain of SD rats Objective to observe and analyze the expression of NG2 + CD15 and nestin in the brain of SD rats at different stages. Methods Sprague-Dawley rats were anesthetized for 7 days and 3 months after birth. The rats were fixed with 4% paraformaldehyde and 10% 20% sucrose after dehydration. According to Wang Pingyu of Xuzhou Medical College, we made frozen sections of the brain of rats and made CD15 on neural stem cell-related antigens (NSC-related antigens). NG2) and O-2A lineage specific marker A2B 5 were studied by immunofluorescence histochemical staining to observe the difference of expression. Results there were almost no nestin and CD15 positive cells in cerebral cortex and medulla in 7-day and 3-month groups, but they were expressed in the vicinity of SVZ and hippocampal dentate gyrus. The expression level decreased slightly with the aging of mice. NG2 was found in different brain regions. The expression of NG2 in mature brain tissues was decreased in subependymal area, but NG2 was expressed in the parenchyma around SVZ area. Conclusion the neural stem cells migrate to the periphery of brain tissue during the development of neural stem cells, and the surface antigen nestin disappears fastest, followed by the slowest or not disappearing during the development of CD15Ng2. The second part: expression of neural stem cell associated antigen in glial cells of O-2A lineage Objective to investigate the expression of NG2 + CD15 and nestin in glial cells of O-2A lineage. Methods according to the difference of growth time and adhesion characteristics of O-2A lineage cells, oscillatory method and differential adhesion method were used. The purified O-2A progenitor cells were obtained by using serum-free medium containing growth factor, and the O-2A progenitor cells were induced to differentiate into T2A by growth factor medium. The O-2A of 1 d and 2 d of culture and T2A of 1 d and 3 d of culture purification were detected by immunofluorescence of nestinine CD15, NG2 and A2B 5, respectively. Results the expression of nestin was stronger in OPC for 1 d and 1 d in the early stage of T2A culture, but decreased slightly (only 70-80%). The expression of CD15 in OPC was stronger than that in T2A, but it was interesting. The expression of CD15 in the differentiated T2A was decreased but not completely unexpressed. NG2 was strongly expressed in the O-2A lineage of OPC T2A. Conclusion during the development of the central nervous system, the expression of neural stem cell markers of O-2A lineage glial cells decreased gradually with the cells from immature to mature. But different neural stem cell markers have different expression patterns.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

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