MTT方法的优化  

The Optimization of MTT Method

作　　者：罗奇志[1] 林琳[1] 王芙艳[1] 余平[1] 

机构地区：[1]中南大学湘雅医学院免疫学系,湖南长沙410008

出　　处：《激光生物学报》2014年第3期279-282,共4页Acta Laser Biology Sinica

基　　金：国家自然科学基金(81371864)

摘　　要：目的：对MTT法检测悬浮细胞增殖活性的实验条件进行筛选。方法：以K562细胞为实验对象，分别测定不同MTT用量、细胞浓度、MTT溶剂种类及作用时间等实验条件下的OD570值。结果：检测K562细胞的增殖活性时，细胞浓度应选取0．8×108～0．2×108/L，MTT加入量不应超过20μL/孔，若不考虑时间成本，应以三联溶液作为甲硂溶剂，反应12 h后检测，所获结果精密度最高；若需快速获得结果，也可选择DMSO作为甲硂溶剂，反应10 min后检测。结论：建立了优化的MTT法检测悬浮细胞增殖活性。Objective：To determine the experimental conditions of MTT method detecting suspension cell proliferation activity.Methods：We took K562 cells as experimental subjects and detected the OD570 value under different experi-mental conditions respectively,including different dosage of MTT,cell concentration,MTT solvent type and reaction time.Results：To detect the proliferation activity of K562 cells,cell concentration should be 0.8×108 ~0.2 ×108/L, the adding amount of MTT should not exceed 20 μL/hole.As time consuming as it was （12 hours after reaction）,the triple-solution was the best formazen solvent for obtaining results with the highest precision,or DMSO could also be a good choice for providing results most quickly （10 minutes after reaction）.Conclusion：We established optimization ex-perimental conditions to detect suspension cell proliferation activity by MTT method.

关 键 词：MTT法 优化 悬浮细胞 

分 类 号：R392-33[医药卫生—免疫学]