恶性胶质瘤细胞株U251中肿瘤干细胞的分离、培养及鉴定

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。

Isolation, Culture and Identification of Brain Cancer Stem Cells from Malignant Glioma Cell Line U251 in Vitro

Aim:To isolate,culture and identificate of brain cancer stem cells from malignant glioma cell line U251 in vitro. Methods:The U251 were cultured in serum-free medium with EGF,bFGF,LIF and B27. The CD133 cells were obtained by magnetic cell sorting from the primary brain tumor spheres,and cultured continuously in the serum-free medium through the monoclonal formation experiment. The immunofluorescence staining was employed to identificate the brain cancer stem cells and the differentiated cells. Results:The brain cancer stem cells from malignant glioma cell line U251 have been successfully isolated. They formed typical neurospheres in serum-free medium,and had the strong capacity to self-renew,and proliferate. The cells expressed CD133 and they could differentiate into neurons and astrocytes. Conclusion:The brain cancer stem cells exist in the malignant glioma cell line U251 in vitro,and they can be isolated,cultured and differentiated.