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目的:从恶性胶质瘤细胞株U251中分离、培养并鉴定肿瘤干细胞。方法:将U251细胞置于含EGF、bFGF、LIF及B27的无血清培养基中培养,形成悬浮生长的细胞球后经免疫磁珠分离获取CD133阳性细胞,采用单细胞克隆法继续在上述培养液中培养。应用细胞免疫荧光染色对肿瘤干细胞及其分化细胞进行鉴定。结果:在恶性胶质瘤细胞株U251中成功分离出肿瘤干细胞,在上述无血清培养液中呈悬浮生长,具有很强的自我更新与繁殖能力,免疫荧光染色显示该细胞表达CD133,诱导分化后可分化成为神经元与星形胶质细胞。结论:体外培养的恶性胶质瘤细胞株U251中存在脑肿瘤干细胞,并能将其分离、培养及诱导分化。
OBJECTIVE: To isolate, culture and identify cancer stem cells from malignant glioma cell line U251. Methods: U251 cells were cultured in serum-free medium containing EGF, bFGF, LIF and B27 to form suspension-growing cytoplasm. CD133-positive cells were isolated by immunomagnetic beads and cultured in the above medium In training. Tumor stem cells and their differentiated cells were identified by immunofluorescence staining. Results: Tumor stem cells were successfully isolated from the malignant glioma cell line U251 and grew in suspension in the above serum-free medium. The cells showed strong ability of self-renewal and reproduction. The immunofluorescent staining showed that the cells expressed CD133 and induced differentiation Differentiation into neurons and astrocytes. Conclusion: There are brain tumor stem cells in U251 cells cultured in vitro, which can be isolated, cultured and induced to differentiate.