利用BD FACSAria Ⅲ二次分选法纯化低比例细胞群体的流式细胞技术  

Flow Cytometry for Purification of Low Proportion Cell Population by BD FACSAria Ⅲ Secondary Separation

作　　者：户乃丽[1] 徐晓雪[1] 邹林樾[1] 赵君朋[1] HU Nai-li;XU Xiao-xue;ZOU Lin-yue;ZHAO Jun-peng(Central Laboratory,Capital Medical University,Beijing 100069)

机构地区：[1]首都医科大学医学中心实验室,北京100069

出　　处：《中国医疗器械信息》2020年第21期19-21,共3页China Medical Device Information

摘　　要：目的:利用BD FACSAriaⅢ流式细胞仪,通过两次分选获得高纯度的GFP阳性HEK293T细胞。方法:GFPpx458空载质粒转染的293T细胞,通过流式细胞仪yield结合purity模式对细胞进行两次分选,比较二次分选后阳性细胞的纯化率与细胞分别经yield和purity单次分选的差异并对分选后细胞进行7-AAD染色鉴定分选后细胞的活性。结论:使用BD FACSAriaⅢ型流式细胞仪的yield加purity模式可将低表达量3.1±1.0%且分群不明显的GFP+293T纯化至96.5±1.1%,且活性良好,是一种可靠的获取高纯度目的细胞的方法。Objective:Using BD FACSAriaⅢflow cytometry sorting for twice to obtain high purity of GFP positive HEK293T cells.Methods:293T cells were transfected with gfp-px458 empty plasmid,the cells were sorted twice by yield combined with purity mode,the purification rate of positive cells after secondary sorting was compared with that of single sorting by yield and purity,and 7-AAD staining was used to identify the activity of the cells.Results:Cell sorting by flow cytometry yield combined with purity mode,GFP+293T with low expression3.1±1.0%and no obvious clustering could be purified to 96.5±1.1%.Cells have good activity.It is a reliable method to obtain high purity target cells.

关 键 词：流式细胞 分选模式 二次分选 

分 类 号：R-331[医药卫生]