TJ905胶质瘤细胞系中脑肿瘤干细胞的培养、分离与鉴定  

Isolation,culture and identification of brain cancer stem cells in glioma cell line TJ905

作　　者：于喜贞[1,2] 韩光魁[2] 杨文涛[2] 李正友[2] 张国安[3] 靳峰[2] 

机构地区：[1]新乡医学院研究生院,新乡市453003 [2]济宁医学院附属医院神经外二科 [3]济宁医学院病理生理学实验室,济宁市272029

出　　处：《广西医学》2015年第1期11-14,共4页Guangxi Medical Journal

基　　金：国家自然科学基金资助项目(8107779);山东省中青年科学家科研奖励基金资助(BS2010YY006)

摘　　要：目的培养恶性胶质瘤TJ905细胞,并从中分离、培养和鉴定肿瘤干细胞,观察干细胞生长特征,为脑肿瘤干细胞的进一步研究奠定基础。方法将TJ905细胞置于含血清培养基中培养;将TJ905细胞置于含生长因子的无血清培养基中培养,待其形成悬浮生长的细胞球后用免疫磁珠分离获取CD133+细胞。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系TJ905中成功分离出肿瘤干细胞,培养于无血清培养液中,细胞呈球形、悬浮生长,具有很强的繁殖与自我更新能力;将该细胞置于含血清的培基中可分化;免疫荧光染色显示脑肿瘤干细胞中神经上皮干细胞蛋白(Nestin)表达阳性。结论培养的恶性胶质瘤细胞系TJ905中有脑肿瘤干细胞的存在,并且能在体外将其培养、分离及诱导分化,可为肿瘤干细胞的研究提供便利条件。Objective To culture malignant glioma cell line TJ905,to isolate, culture and identify the brain cancer stem ceils, and to observe the features of stem cell growth, aiming at laying the foundation for the further study of brain tumor stem ceils. Methods The TJg05 ceils were cultured in serum medium. After the TJ905 cells were cultured in serum-free medium with growth factor, the CD133 ＋ cells were isolated and obtained by magnetic cell sorting from the suspended growth ceil spheres. The staining was adopted to identify the brain cancer stem cells and their differentiated ceils. Results The cancer stem cells were isolated from malignant glioma cell line TJ905 successfully. The cells cultured in serum-free medium were spherical, with suspension ceil growth pattern,which had the strong capacity of proliferation and self-renewal. The cells could be differentiated when they were cultured in the serum medium. The result of immunofluorescence staining showed a positive expression of the neuroepithelial stem cell protein （Nestin） in brain cancer stem cells. Conclusion The cancer stem ceils exist in the malignant glioma cell line TJ905,and they can be cultured,isolated and differentiated in vitro, which might facilitate the study of cancer stem ceils.

关 键 词：肿瘤干细胞 TJ905细胞 胶质瘤 免疫磁珠细胞分选 分离 培养 鉴定 

分 类 号：R739.41[医药卫生—肿瘤]