胶质母细胞瘤原代细胞单克隆株的建立  

Establishment of single cell colony from human glioblastoma

作　　者：周强[1] 漆松涛[1] 向伟[1] 王海[1] 张立[1] 李宏[1] 刘亚伟[1] 陆云涛[1] 

机构地区：[1]南方医科大学南方医院神经外科,广州510515

出　　处：《中华实验外科杂志》2015年第9期2074-2076,共3页Chinese Journal of Experimental Surgery

基　　金：广东省自然科学基金资助项目（2014A030313298）;南方医科大学南方医院院长基金资助项目（2014Z002）

摘　　要：目的 建立胶质母细胞瘤原代细胞的单克隆株并观察记录不同单克隆株间细胞形态和增殖能力的差异.方法 胶质母细胞瘤组织原代培养后进行纯化、单克隆培养、鉴定;定期观察单克隆细胞株的生长状况和形态特征,并比较其增殖能力的差异.结果 成功建立了23株胶质母细胞瘤细胞单克隆株,根据单克隆株形态不同,将其分为长伪足型和短伪足型,长伪足型伪足长（142.50±33.65） μm,短伪足型伪足长（31.33 ±5.72） μm,两者伪足长短差异有统计学意义（P＜0.01）;同时,细胞增殖实验证明长伪足型单克隆细胞株增殖能力弱于短伪足型单克隆细胞株,细胞计数试剂盒（CCK--8）实验显示长伪足型单克隆株第10天吸光度值为0.85 ±0.13,短伪足型单克隆株吸光度值为1.31 ±0.20,两者差异有统计学意义（P＜0.01）.结论 不同单克隆细胞株形态和增殖能力的差异印证了肿瘤细胞的异质性.Objective To establish single cell colony from human glioblastoma multiforme （GBM） and observe corresponding morphological characteristics and proliferation capacity.Methods GBM cells were acquired by enzymatic digestion culture method and further purified for single clone culture.Subsequently,these glial fibrillary acidic protein （GFAP） positive colonies were indentified for research.At last,cell growth and morphological characteristics of single cell colony were recorded.Results We successfully establish 23 single cell colonies and divide them into short pseudopod type and long pseudopod type according to their morphological diversity,the length of pseudopod is （142.50 ± 33.65） μm in long pseudopod group and （31.33 ± 5.72） μm in short pseudopod group respectively,length of these two type cell are significantly different （P ＜ 0.01）.In addition,cell counting kit-8 （CCK-8） assay indicate short pseudopod type cell exhibit significant higher proliferation capacity than long pseudopod type cell.The OD value at 10th day is 0.85 ±0.13 in long pseudopod group and 1.31 ±0.20 in short pseudopod group respectively （P ＜ 0.01）.Conclusion Diversity of morphology and proliferation capacity among cell colonies partially demonstrate existing of intratumor heterogeneity.

关 键 词：胶质母细胞瘤 原代培养 单克隆 有限稀释法 

分 类 号：R730.264[医药卫生—肿瘤]