神经胶质细胞的干细胞抗原表型研究
发布时间：2018-10-24 19:22
【摘要】：目的检测新生大鼠大脑皮质来源的星形胶质细胞及小胶质细胞的干细胞抗原表达,为进一步深入研究这两种胶质细胞的应用意义提供生物学实验依据。 方法1.取新生大鼠大脑皮质进行原代混合胶质细胞培养,根据各种胶质细胞生长时间差异及细胞的粘附特性不同,通过机械震荡和差速贴壁方法分离纯化O-2A祖细胞、小胶质细胞(microglia, MGs)和1型星形胶质细胞(type 1 astrocytes, T1As),使用细胞特异性抗体进行鉴定。 2.收集O-2A祖细胞接种于12孔培养板或/和包被PLL的盖玻片上,培液为DMEM/F-12、2% B27、1% N2、10ng/ml PDGF,进一步纯化O-2A祖细胞。在含20%FCS的培养条件下诱导O-2A祖细胞向2型星形胶质细胞(type 2 astrocytes, T2As)分化,使用细胞特异性抗体进行鉴定。 3.采用免疫荧光标记染色方法,检测干细胞的标记分子巢蛋白(nestin)、阶段特异性胚胎抗原-1(SSEA-1)及蛋白聚糖NG2在星形胶质细胞和小胶质细胞的表达情况。 结果1.通过机械震荡和差速贴壁方法,有效分离了O-2A祖细胞及1型星形胶质细胞,它们具有典型的形态特征,表达各自的细胞特异性分子,O-2A祖细胞表达A2B5,1型星形胶质细胞表达GFAP。 2.通过机械震荡和差速贴壁方法,有效分离了纯化的小胶质细胞,它们具有典型的形态特征,并表达其细胞特异性分子OX42。 3.分离的O-2A祖细胞经PDGF作用,增殖而不出现分化,从而O-2A祖细胞得到纯化。 4.O-2A祖细胞以无血清培养液培养,迅速分化为成熟的少突胶质细胞；在含高达15%-20%FCS的培液中生长,分化为2型星形胶质细胞,特异性抗原表达为A2B5+/GFAP+。 5.免疫荧光标记染色显示：分化成熟的2型星形胶质细胞表达nestin, SSEA-1及NG2,小胶质细胞表达nestin, SSEA-1及NG2,1型星形胶质细胞不表达干细胞抗原标志物。 结论神经干细胞标记分子nestin, SSEA-1及NG2在胶质细胞中的表达存在差异。2型星形胶质细胞及小胶质细胞均表达nestin, SSEA-1及NG2,具有干细胞的某些抗原表达特征,结合我研究室已有研究表达提示,2型星形胶质细胞及小胶质细胞具有神经干细胞样的生物学特性,可作为细胞来源应用于中枢神经系统疾病的治疗研究。
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【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329
本文编号：2292321
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【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329
【参考文献】
相关期刊论文 前2条
1 张晔;李春鹏;夏春林;沈慧;孙文阁;;蛋白脂蛋白在两型星形胶质细胞和少突胶质细胞中的表达[J];解剖学杂志;2006年05期
2 段树民;;胶质细胞的细胞间通讯[J];生命科学;2008年05期
，本文编号：2292321
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