流式分选仪FACSAriaⅡ分选GFP阳性乳腺癌MCF-7细胞株的条件优化研究

发表时间：2014-06-23 浏览次数：1281次

引用：

郭陈智，史桂英.流式分选仪FACSAriaⅡ分选GFP阳性乳腺癌MCF-7细胞株的条件优化研究[J].上海交通大学学报（医学版）2013, 33(1).127-

关键词：

作者：

郭陈智，史桂英

作者单位：

(上海交通大学基础医学院生物化学与分子细胞生物学系公共平台流式实验室,上海,200025)

出版年份：

2013

期刊页数：

127-130

收录者：

知网,万方

摘要：

目的研究流式分选仪FACSAriaⅡ分选绿色荧光蛋白(GFP)阳性乳腺癌MCF-7细胞株时不同条件选择对分选后细胞的纯度、得率和活性的影响.方法流式分选仪FACSAriaⅡ分别选择70、85和100 μm三种喷嘴以及4-way purity、purity、yield三种分选模式分选MCF-7细胞的GFP阳性稳转株,计数分选后的细胞并计算分选得率,流式细胞术检测分选细胞的纯度,锥虫蓝染色法检测分选细胞的活性.结果 70、85和100 μm三种喷嘴分选GFP阳性细胞纯度均达99％以上,分选得率分别为(37.67±2.05)％、(56.33±2.62)％和(43.33±0.47)％,活细胞比例分别为(88.23±0.97)％、(91.87±1.05)％和(95.33±0.90)％；选择85 μm喷嘴,4-way purity、purity和yield三种分选模式分选细胞的得率分别为(51.00±2.45)％、(67.33±2.05)％和(74.00 ±0.82)％,纯度分别为99.6％、99.3％和96.2％.结论流式分选仪FACSAriaⅡ分选乳腺癌MCF-7细胞GFP阳性稳转株,选择85 μm喷嘴以及purity分选模式较合适. Objective To sort green fluorescent protein(GFP) positive population of breast cancer cell line MCF-7 by flow cytometry sorter FACSAria II，and investigate the effect of different setup on the purity, yield and viability of collected cells. Methods GFP positive population of MCF-7 cells was sorted by 70 p,m, 85 pm and 100 p.m nozzles and 4-way purity，purity and yield sort modes respectively. The yield was calculated with cell counting, the purity was analyzed by flow cytometry, and the viability was determined by trypan blue staining. Results FACSAria ]j gained purity of more than 99% with all three nozzles. The yields of 70协m, 85 p,m and 100协m nozzles were (37. 67士2. OS)%，(56. 33士2. 62)% and (43. 33士0. 47 )%respectively, while the percentages of live cells were(88. 23士0. 97 )%，( 91. 87士1. OS)%and (95. 33 10. 90)%respectively. Sorting cells by 4-way purity, purity and yield sort modes with 85 N,m nozzle, the yields were ( 51. 00士2. 45)%，( 67. 33士2. OS)%and(74. 00士0. 82)%respectively, and the purities were 99.6%，99.3 % and 96.2% respectively. Conclusion In sorting GFP positive population of breast cancer cell line MCF-7 by FACSAria n，85 p,m nozzle and purity sort mode may be the favorable setup.

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