人脑胶质瘤相关间充质干细胞的分离、鉴定及生物特性研究

Isolation, identification and biological characteristics of glioblastoma-associated mesenchymal stem cells

目的 从人脑胶质瘤新鲜标本中分离相关间充质干细胞完成鉴定并对其生物特性进行初步研究.方法 将肿瘤组织经过机械法和胰酶消化后经梯度离心法所得的单个细胞悬液使用磁珠分离法筛选CD133阴性细胞进行培养.使用流式细胞术分析表面标志物,使用细胞计数法分析细胞亚群的生长曲线和促肿瘤细胞增殖作用.结果 脑胶质瘤相关间充质干细胞呈贴壁生长,该细胞阳性表达CD73、CD90、CD105,而阴性表达CD14、CD45、CD31,体外诱导条件下可向脂肪细胞和成骨细胞分化,且其条件培养液可促进脑胶质瘤细胞的增殖.结论 胶质瘤相关间充质干细胞存在于脑胶质瘤微环境中,并可能对脑胶质瘤的发展起促进作用.

Objective To isolate glioblastoma-associated mesenchymal stem cells （gbMSCs） from fresh human glioblastoma tissues,and investigate the identification and biological characteristics.Methods A cell population was isolated from glioblastoma tissues and was defined as gbMSCs by fluorescence-activated cell sorter （FACS） and differentiation assay.The growth curve and the promotion of gbMSCs were analyzed by cell counting.Results GbMSCs were plastic-adherent cells under standard culture condition;these cells have positive expression of CD105,CD90,CD73,and negative expression of CD14,CD31,CD45; In vitro,mesenchymal stem cells can differentiate into osteocytes,adipocytes under specific stimuli.Moreover,the conditioned medium could promote the proliferation of glioma cells.Conclusion gbMSCs could be found in the tumor microenvironment and may promote the progression of glioblastoma.