论文部分内容阅读
目的分离、培养和鉴定胶质瘤中脑肿瘤干细胞(BTSC),探讨CD133作为BTSC特异性标记,用于BTSC分离的可行性。方法9例胶质瘤临床标本,胶质瘤C6和U87细胞系,用直接培养和磁珠选择CD133+细胞二种方法分离BTSC。参照NSC培养条件,进行体外培养。观察细胞生长情况,利用免疫荧光染色检测细胞标记,包括Nestin,神经元特异性烯醇化酶(NSE),胶质纤维酸性蛋白(GFAP)等;诱导细胞分化,验证该种细胞是否具有干细胞特性。结果9例临床标本中,有2例儿童恶性胶质瘤在培养过程中出现神经球,在成人胶质瘤中均未出现。C6和U87细胞中,均出现比较典型的神经球,二种分离方法结果相同。神经球细胞具有增殖能力,体外培养可以不断增大。标记检测显示这种细胞Nestin阳性,而NSE和GFAP阴性。经诱导后,可以分化为形态与来源细胞相似的细胞,能够表达NSE和GFAP等多种表型细胞标记。结论在胶质瘤中,存在具有NSC特性的干细胞,即BTSC。CD133是BTSC重要的表面标记,可以用于BTSC的分离。在胶质瘤细胞系中CD133+细胞是BTSC。
Objective To isolate, culture and identify human glioma meningioma stem cells (BTSCs) and to explore the feasibility of using CD133 as a BTSC-specific marker for BTSC isolation. Methods Nine glioma specimens, C6 and U87 glioma cell lines, were isolated by direct culture and magnetic bead selection of CD133 + cells. Reference NSC culture conditions, in vitro culture. Cell growth was observed. Cell markers such as Nestin, NSE and GFAP were detected by immunofluorescence staining. Cells were induced to differentiate to determine whether the cells had stem cell characteristics. Results Of 9 clinical samples, 2 cases of malignant gliomas in children showed neurospheres during culture, and none appeared in adult gliomas. C6 and U87 cells, there are more typical neurospheres, the results of the two methods of separation of the same. Neurosphere cells have proliferative capacity, in vitro culture can continue to increase. Marker assays revealed Nestin-positive cells, while NSE and GFAP-negative cells. After induction, it can differentiate into cells with similar morphology and origin, and can express many phenotypic cell markers such as NSE and GFAP. Conclusion In gliomas, stem cells with NSC characteristics exist, namely BTSC. CD133 is an important surface marker for BTSC and can be used for BTSC separation. CD133 + cells are BTSCs in glioma cell lines.