贵州医科大学学报

2017, v.42;No.206(11) 1253-1257

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流式实验试剂的使用和操作对贴壁细胞的细胞周期结果的影响
Effect of Laboratory Reagent Application and Practice on Flow Cytometry Tested Adherent Cells' Cell Cycle

周艳华;黄明琦;范安然;舒莉萍;刘杰麟;何志旭;
ZHOU Yanhua;HUANG Mingqi;FAN Anran;SHU Liping;LIU Jielin;HE Zhixu;Stem Cell and Tissue Engineering Research Center;College of Food Safety,Guizhou Medical University;

摘要(Abstract):

目的:分析实验试剂的使用和操作对流式细胞术检测贴壁细胞的细胞周期结果的影响。方法:根据在染色液中是否添加RNA酶、收集贴壁细胞时是否收集培养液及操作时是否吹打的方式,将混悬细胞分为6组(每种处理方式分2组),用碘化丙啶(PI)试剂盒对MBA-MD-231细胞进行染色,采用FC-500流式细胞仪对贴壁细胞的细胞周期进行检测,观察染色液中是否添加RNA酶、是否收集培养液、或是否吹打对细胞流式信号分布图的影响。结果:染色液中添加RNA酶会造成流式信号分布图中细胞周期检测信号峰度变窄,各时期细胞比例发生变化;与仅收集贴壁细胞组相比,收集培养液时细胞碎片百分比略有增加(2.18%vs 1.24%),但对细胞周期检测结果没有影响;混悬细胞时过度吹打(≥30次)不影响细胞周期检测结果,但会造成细胞碎片百分比略有增加(2.31%vs1.24%)。结论:采用流式细胞术分析细胞周期时须添加RNA酶,实验操作时是否收集培养液和吹打细胞对细胞周期检测结果的影响不大。
Objective: To investigate the effect of laboratory reagent application and practice on Flow Cytometry tested adherent cells' cell cycle. Method: Dividing suspension cells into 6 groups( 2 groups for each treatment method) according to whether adding RNA enzyme in staining fluid,collecting culture solution while collecting adherent cells and blowing or beating while practicing or not; MBA-MD-231 cells were stained with propidium iodide( PI) kit,and the cell cycle was detected by FC-500 flow cytometry. Observing staining fluid was added with RNA enzyme or not,collecting culture solution,blowing or beating while practicing affected cell streaming signal distribution. Results: Staining adding RNA enzymes lead cell cycle's signal peak narrowed,proportion of different phases of cells changed;cell debris rates were slightly increased while all culture medium was collected compared with only collecting adherent cells collected( 2. 18% vs 1. 24%),which showed no influence on cell cycle test; over blowing and beating during suspension solution( ≥30 times) would not affect cell cycle test result,but it would slightly increase cell debris rate( 2. 31% vs 1. 24%). Conclusion: RNA enzyme is necessary when analyzing the cell cycle by flow cytometry; collecting culturing solution or blowing and beating suspension solution or not had little effect on the results.

关键词(KeyWords): 贴壁细胞;细胞周期;流式细胞术;细胞碎片
adherent cells;cell cycle;flow cytometry;cell debris

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(31660316);; 贵州省应用基础研究计划重大专项[黔科合J重大字(2015)2003]

作者(Author): 周艳华;黄明琦;范安然;舒莉萍;刘杰麟;何志旭;
ZHOU Yanhua;HUANG Mingqi;FAN Anran;SHU Liping;LIU Jielin;HE Zhixu;Stem Cell and Tissue Engineering Research Center;College of Food Safety,Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2017.11.003

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