流式细胞仪、免疫磁珠及亲和板结合分离法分选c-kit^＋肝癌细胞的比较  

Comparative study of isolating c-kit-positive hepatoma carcinoma cells with flow cytometry sorting, magnetic cell sorting and immune panning isolation

作　　者：刘胜军[1] 方驰华[1] 

机构地区：[1]南方医科大学珠江医院肝胆外科,广州510282

出　　处：《中华消化外科杂志》2007年第3期206-208,共3页Chinese Journal of Digestive Surgery

基　　金：广东省自然科学基金资助项目（2001010593：2002020097）

摘　　要：目的比较流式细胞仪、免疫磁珠及亲和板结合分离法分选肝癌亚群细胞的效果。方法原代培养肝细胞癌细胞,分别以流式细胞仪、免疫磁珠及亲和板结合分离法分选c-kit^＋肝癌细胞,然后使用流式细胞仪鉴定细胞纯度,计算回收率;以台盼蓝检测细胞活力,CCK-8法检测细胞增殖能力,裸鼠移植测其成瘤率,并进行统计学分析。结果流式细胞仪分选所得到的c-kit^＋肝癌细胞纯度、回收率最高,与其他两种方法分选所得细胞的相应指标比较,差异有统计学意义（P〈0.05）;3种方法所得细胞之间的细胞活力、刺激指数、移植成瘤率比较,差异无统计学意义（P〉0.05）。流式细胞仪分选前后的细胞之间活力、刺激指数比较,差异无统计学意义（P〉0.05）。结论流式细胞仪分离法是分离肝癌亚群细胞的较佳方法。Objective To compare the effect of three methods including flow cytometry sorting, magnetic cell sorting and immune panning isolation in isolating sub - population in hepatoma carcinoma cells. Methods c-kit-positive cells in primarily cultured human hepatoma carcinoma cells were separated with flow cytometry sorting, magnetic cell sorting and immune panning isolation, respectively. The purity, viabililty, cell proliferation and the ratio of tumorigenesis after transplantation of the hepatoma carcinoma cells were determined with flow cytometry, trypan blue exclusion, CCK-8 assay and allogeneic graft to nude mouse respectively. Then, the data were compared statistically. Results The purity and recovery rate of the c-kit-positive cells isolated with flow cytometry sorting were significantly higher than those with the other two methods, with statistical difference （ P 〈 0. 05 ）. There was no significant difference upon viablihy, stimulation index and tumorigenesis after transplantation among the cells isolated with the three methods （ P 〉 0. 05 ）. There was no significant difference upon viability and proliferation between the cells before and after separating by flow cytometry sorting （ P 〉 0.05 ）. Conclusions Flow cytometry sorting is an effective and practical method for isolating sub-population in hepatoma carcinoma cells.

关 键 词：流式细胞仪 免疫磁珠 亲和板结合分离法 

分 类 号：R686[医药卫生—骨科学]