CD133免疫磁珠分选脑肿瘤干细胞及其生物学特性  

CD133 magnetic sorting and biological characteristics of brain tumor stem cells

作　　者：齐玲[1] 金宏[1] 丁丽娟[2] 温娜[1] 唐泽波[1] 狄阳[1] 李蕴潜[2] 

机构地区：[1]吉林医药学院病理学教研室,吉林吉林132013 [2]吉林大学第一医院神经外科,吉林长春130021

出　　处：《吉林大学学报（医学版）》2011年第3期441-444,I0002,共5页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅自然科学基金资助课题(202015242);吉林省教育厅科研基金资助课题(吉教科合字[2011]第281号;吉教科合字[2010]第532号)

摘　　要：目的:从人脑肿瘤组织中分离、培养、纯化和鉴定脑肿瘤干细胞(BTSCs),探讨CD133免疫磁珠分选方法获取BTSCs的可行性及BTSCs的生物学特性。方法:留取人脑胶质母细胞瘤新鲜标本,分离获得脑肿瘤细胞。应用CD133免疫磁珠分选方法纯化BTSCs;流式细胞仪检测分选阳性率;神经球计数法分析CD133+/-亚群细胞神经球形成情况;免疫荧光方法检测CD133+/-亚群细胞表面神经干细胞(NSCs)标记物CD133、神经巢蛋白(nestin)表达情况;检测CD133+/-亚群细胞经诱导分化后细胞表面分化标记物β-TubulinⅢ、GFAP表达情况。结果:CD133+亚群细胞具有干细胞特性,可形成明显的神经球,具有自我更新和明显的增殖能力,并且NSCs标记物CD133、nestin表达阳性,经诱导分化后分化标记物β-TubulinⅢ、GFAP表达阳性;CD133-亚群细胞无上述特性。结论:CD133免疫磁珠分选方法获得的CD133+亚群细胞即为BTSCs,该方法可以得到高纯度的BTSCs,可以用于BTSCs的实验研究。Objective To isolate and culture brain tumor stem cells（BTSCs）,and explore the feasibility for obtaining BTSCs by CD133 magnetic sorting and the biological characteristics of BTSCs.Methods The fresh glioblastoma sample was dissociated,the cells were cultivated in serum-free media with growth factor,the BTSCs were purified by CD133 magnetic sorting,the positive rate of CD133＋/-subgroup cells was detected by flow cytometry;neurosphere forming essay was used to analyze the self-renewal and proliferative capacity;immunofluorescent staining was performed to detect the neural stem cells（NSCs） markers CD133 and nestin of CD133＋/-subgroup cells,and the multipotentiality markers β-TubulinⅢ and GFAP of CD133＋/-subgroup cells after induction.Results The CD133＋ cells had the characteristics of stem cells,and could form the neurosphere,had the self-renewal and proliferative capacity.The expressions of CD133 and nestin were positive in CD133＋ cells;β-TubulinⅢ and GFAP were also positive after differentiation.But CD133-cells hadn＇t the characteristics mentioned above.Conclusion CD133 magnetic sorting can isolate and purify BTSCs,it can be used to get more BTSCs for experiments.

关 键 词：脑肿瘤干细胞 神经干细胞 CD133 神经巢蛋白 

分 类 号：Q813[生物学—生物工程] R739.41[医药卫生—肿瘤]