胶质瘤干细胞的分离、培养及初步鉴定

摘要

Objective:To explore the culture conditions for glioma stem cells(GSCs)and to investigate the differ-entiation potential of GSCs.Methods:The glioma tissues from 9 patients were used to generate GSCs.Cells were cul-tured in B27 medium with basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF),and they were i-dentified by immunocytochemistry.Results:CD133 -positive GSCs were successfully cultured from 7 out 9 patients. They formed typical neurospheres in suspension,and could be cultured and passaged steadily.The majorities of the cells expressed CD133 and Nestin,which were the markers for neural stem cells,and expressed GFAP and TU -20 respectively.Conclusion:GSCs could be cultured from gliomas in vitro,which provided a basis for further studies.%目的：探讨人脑胶质瘤干细胞的培养和分离方法。方法：取9例脑胶质瘤患者的手术标本进行处理，采用 B27培养基进行培养，碱性成纤维细胞生长因子和表皮生长因子刺激细胞扩增；应用免疫组织化学染色对培养的细胞及其分化的细胞进行鉴定。结果：9例标本中共有7例成功培养出肿瘤细胞球，培养条件下呈悬浮状态生长，形成肿瘤细胞球，免疫细胞化学检测显示肿瘤球细胞表达胶质瘤干细胞的标志物 CD133和 Nestin，诱导分化后的肿瘤球细胞可以表达成熟神经细胞的标志物 GFAP 和 TU －20。结论：体外的培养条件下，可以从胶质瘤组织中培养出胶质瘤干细胞，为胶质瘤的深入研究奠定基础。