免疫磁珠分选系统在分离肝癌干细胞中的应用

Application of magnetic activated cell sorting system in the isolation of liver cancer stem cells

目的为进一步研究肝癌干细胞的特性,应用免疫磁珠分选系统分离纯化肝癌细胞系Huh-7中的肝癌干细胞。方法采用EpCAM单克隆抗体包被的微小磁珠(平均直径≤50 nm)标记Huh-7细胞,通过免疫磁珠分选系统分离EpCAM(epithelial cell adhesion molecule,EpCAM)阳性的肝癌干细胞,以流式细胞术检测分离后的肝癌干细胞纯度,以台盼蓝染色法检测细胞活性。结果采用流式细胞仪检测分离前肝癌细胞中EpCAM阳性细胞比率为(13.3±1.106)%,分离后的肝癌干细胞中EpCAM阳性细胞比率达到(99.5±0.200)%,细胞存活率为(96.5±1.518)%。结论采用免疫磁珠分选系统分离肝癌干细胞操作简便、快速有效,为肝癌干细胞的进一步研究奠定了基础。

Objective To isolate the liver cancer stem ceils from the hepatocellular carcinoma cell line Huh-7 by magnetic activated cell sorting system （MACS） for further studying the property of these ceils. Methods Huh-7 Cells were labeled by EpCAM monoclonal antibody coated with micro beads（mean diameter ≤50 nm）. EpCAM positive liver cancer cells were separated by MACS. Purity of the sorted cells was determined by flow cytometry. The viable sorted cells were counted by Trypan blue staining. Results The rate of EpCAM positive cells was（13.3±1.106）% in Huh-7 cells by flow cytometry analysis before sorting. Rate of EpCAM positive cells was （99.5±0.200）% in the sorted cells. The percent of viable sorted cells was（96.5±1.518）%. Conclusion The magnetic cell sorting system could separate liver cancer stem cells easily,rapidly and effectively from hepatocellular carcinoma cells,which lays foundation for further studying.