恶性胶质瘤细胞株U251中肿瘤干细胞的分离、培养及鉴定  

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

作　　者：王彬[1] 杨辉[1] 尹昌林[1] 吕胜青[1] 刘海鹏[1] 

机构地区：[1]第三军医大学附属新桥医院神经外科,400037

出　　处：《中国临床神经科学》2007年第6期638-642,共5页Chinese Journal of Clinical Neurosciences

摘　　要：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。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.

关 键 词：胶质瘤 脑肿瘤干细胞 培养 免疫磁珠 

分 类 号：Q198[生物学—普通生物学]