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目的:探讨脑肿瘤干细胞(brain tumor stem cells,BTSCs)的分离、培养及鉴定的方法,研究BTSCs比例与人脑神经上皮肿瘤病理级别之间的相关性。方法:将人神经上皮肿瘤细胞接种于含生长因子的无血清培养基中,使其中的脑肿瘤干细胞增殖和传代,利用细胞免疫荧光及免疫组化法检测干细胞的特异性标记物Nestin和CD133的表达;采用免疫组化(SABC法)研究46例脑肿瘤及5例正常脑组织CD133的表达,并与肿瘤病理分级进行相关性分析。结果:可以从人神经上皮肿瘤病例中分离培养出BTSCs;BTSCs可以在体外增殖并传代,并且表达干细胞的特异性标记物Nestin和CD133;CD133在正常的脑组织中未见表达,在各级神经上皮肿瘤中均有表达,各级别肿瘤中的阳性细胞比例差异有显著性(P<0.01),CD133阳性细胞比例与肿瘤的恶性程度呈正相关(P<0.01)。结论:各种病理类型神经上皮肿瘤组织中均存在干细胞特征的细胞,CD133的表达与肿瘤级别呈正相关。
OBJECTIVE: To investigate the isolation, culture and identification of brain tumor stem cells (BTSCs) and to study the correlation between the proportion of BTSCs and the pathological grade of human brain epithelial neoplasia. Methods: Human neural epithelial tumor cells were seeded in serum-free medium containing growth factors to proliferate and pass the brain tumor stem cells. Immunofluorescence and immunohistochemistry were used to detect the expression of Nestin and CD133 The expression of CD133 in 46 cases of brain tumor and 5 cases of normal brain tissue was studied by immunohistochemistry (SABC method), and the correlation was analyzed with the pathological grade of tumor. RESULTS: BTSCs could be isolated and cultured from human neuroepithelial neoplasia cases. BTSCs proliferated and passaged in vitro, and expressed stemin-specific markers Nestin and CD133. CD133 was not expressed in normal brain tissues. (P <0.01). The positive rate of CD133 positive cells was positively correlated with the malignant degree of tumor (P <0.01). CONCLUSIONS: Stem cells are present in all pathological types of neuroepithelial neoplasms. CD133 expression is positively correlated with tumor grade.