上海交通大学学报（医学版）

›› 2013, Vol. 33 ›› Issue (1): 127-.doi: 10.3969/j.issn.1674-8115.2013.01.026

• 技术与方法 • 上一篇    

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

郭陈智, 史桂英   

  1. 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
  • 出版日期:2013-01-28 发布日期:2013-02-06
  • 作者简介:郭陈智（1985—）, 男, 助理实验师, 硕士；电子信箱: guochenzhi@gmail.com。

Study on favorable setup of flow cytometry sorter FACSAriaⅡ in sorting GFP positive population of breast cancer cell line MCF-7

GUO Chen-zhi, SHI Gui-ying   

  1. Flow Cytometry Facility, Department of Biochemistry and Molecular Cell Biology, Basic Medical College, Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2013-01-28 Published:2013-02-06

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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。

关键词: 流式分选, 喷嘴, 分选模式, 纯度, 得率

Abstract:

Objective To sort green fluorescent protein (GFP) positive population of breast cancer cell line MCF-7 by flow cytometry sorter FACSAriaⅡ, 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 μm, 85 μm and 100 μm 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 FACSAriaⅡ gained purity of more than 99% with all three nozzles. The yields of 70 μm, 85 μm and 100 μm nozzles were (37.67±2.05)%, (56.33±2.62)% and (43.33±0.47)% respectively, while the percentages of live cells were (88.23±0.97)%, (91.87±1.05)% and (95.33±0.90)% respectively. Sorting cells by 4-way purity, purity and yield sort modes with 85 μm nozzle, the yields were (51.00±2.45)%, (67.33±2.05)% and (74.00±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 FACSAriaⅡ, 85 μm nozzle and purity sort mode may be the favorable setup.

Key words: cytometry sorting, nozzle, sort mode, purity, yield