C6胶质瘤细胞系中脑肿瘤干细胞的悬浮培养和鉴定  

The Suspension Culture and Identification of Brain Tumor Stem Cells from C6 Glioma Cell Line

作　　者：冯利强[1,2] 朱艳[3] 张自新[4] 刘印明[1,2] 马江波[1,2] 丁银秀[1,2] 

机构地区：[1]宁夏医科大学颅脑疾病重点实验室,银川750004 [2]宁夏医科大学基础医学院人体解剖和组织胚胎学系,银川750004 [3]宁夏回族自治区人民医院(西北民族大学第一附属医院)宁夏眼科医院,银川750004 [4]宁夏医科大学总医院肿瘤医院放疗科,银川750004

出　　处：《宁夏医科大学学报》2017年第1期24-26,37,F0004,共5页Journal of Ningxia Medical University

基　　金：宁夏自然科学基金(NZ13277);宁夏医科大学校级课题(XZ201423);国家自然科学基金地区项目(81360062)

摘　　要：目的观察C6胶质瘤细胞系中脑肿瘤干细胞的悬浮培养、增殖、分化的生物学特征。方法进行C6BTSCs的悬浮培养,应用倒置相差显微镜观察干细胞的生长特性;免疫荧光染色观察Brd U、CD133和Nestin的表达情况。结果在无血清条件培养基中,悬浮培养的C6 BTSCs增殖速度快,具有较强的增殖分裂能力,呈BrdU、CD133和Nestin强阳性,其中Brd U阳性细胞数占CD133阳性细胞数的65%以上;在含血清培养基中,BTSCs呈神经样细胞分化,其细胞形态与贴壁培养的C6胶质瘤细胞在形态上无差别。结论 C6胶质瘤细胞系中存在脑肿瘤干细胞,且该细胞可悬浮培养、增殖和分化。Objective To investigate the suspension culture,proliferation and differentiation of C6 brain tumor stem cells isolated from C6 glioma cell line in vitro. Methods C6 brain tumor stem cells were suspensing cultured in vitro and cells growth observed by phase contrast microscope. Mixed with Brd U in culture medium,the expressions of Brd U,CD133 and Nestin on C6 brain tumor stem cells was identified by immunocytochemistry. Results Under the function of conditioned free-serum culture medium,brain tumor stem cells grew in suspension of cell spheres,with higher speed and much more stem cells. Proceeding successive passage,the biological property of differentiated C6 brain tumor stem cells is same with primary C6 glioma cell line. Conclusion Brain tumor stem cells exist in the C6 glioma cell line,which can be suspensed culture,and have characteristics of proliferation and differentiation in vitro. The discovery of brain tumor stem cells provides a new way for targeted therapy of brain glioma diseases.

关 键 词：C6脑肿瘤干细胞 悬浮培养 增殖 BrdU掺入实验 

分 类 号：R739.41[医药卫生—肿瘤] Q813[医药卫生—临床医学]