恶性胶质瘤中干细胞分离与细胞周期检测  

Isolation of stem cells from malignant glioma and detection of the cell cycle

作　　者：王彬[1] 杨辉[1] 尹昌林[1] 吕胜青[1] 刘海鹏[1] 黄其林[1] 

机构地区：[1]第三军医大学附属新桥医院神经外科,重庆400037

出　　处：《中华神经外科疾病研究杂志》2009年第5期405-408,共4页Chinese Journal of Neurosurgical Disease Research

基　　金：第三军医大学青年创新基金资助项目(2007XG30)

摘　　要：目的从多形性胶质母细胞瘤组织标本中分离、培养、鉴定胶质瘤干细胞,并检测其所处的细胞周期。方法采用逐步递减培养基中血清含量的方式从多形性胶质母细胞瘤组织标本中获得悬浮生长的肿瘤球,应用免疫荧光染色检测胶质瘤干细胞及其分化细胞表面标志物的表达,免疫组化检测裸鼠颅内移植瘤表型。免疫磁珠分选多形性胶质母细胞瘤组织标本中的CD133阳性细胞后立即检测细胞周期。结果在多形性胶质母细胞瘤组织标本中成功分离出胶质瘤干细胞,在无血清培养液中呈悬浮生长,具有很强的自我更新与繁殖能力,免疫荧光染色显示该细胞表达CD133和巢蛋白,诱导分化后可分化成为神经元、星形胶质细胞与少突胶质细胞,裸鼠颅内移植后可重现亲本肿瘤表型。位于其中的胶质瘤干细胞大多处于G0～G1期。结论多形性胶质母细胞瘤组织中存在胶质瘤干细胞,相对处于静止状态。Objective To isolate, culture and identify stem cells from glioblastoma multiforme and detect the cell cycle. Methods The suspending growing neurospheres from glioblastoma multiforme were harvested through gradually reducing serum concentration. The immunofluorescence staining was employed to identify the brain glioma stem cells and their differentiated cells. The immunochemistry staining was used to identify the transplanted brain tumor and detect the cell cycle of CD133 ^＋ after magnetic cell sorting. Results The brain glioma stem cells from glioblastoma multiforme had been successfully isolated. They formed typical neurospheres in serum-free medium with the strong capacity of self-renewing and proliferating. The cells expressed both CD133 and nestin in immunofluorescence staining and they could also differentiate into multi-lineage progenies. The cells could produce brain tumors in NOD-SCID mice and the tumors were the phenocopy of the original tumor from which they were derived. The brain glioma stem cells in glioblastoms maltiforme were in G0-G1 phase. Conclusion The brain glioma stem cells do exist in glioblastoma multiforme at an inactive mitotic division status.

关 键 词：多形性胶质母细胞瘤 胶质瘤干细胞 培养 细胞周期 

分 类 号：R739.41[医药卫生—肿瘤]