C6胶质瘤细胞系中脑肿瘤干细胞的悬浮培养和鉴定

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BrdU、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BrdU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 BrdU in culture medium,the expressions of BrdU,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.