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本实验用2种体外传代培养的白血病细胞株(K_(562),HL_(60))对MTT实验条件进行研究。结果表明:MTT代谢产物甲 分细胞数量呈线性关系。最佳实验条件为:实验用细胞数为2×10~5/孔或4×10~5/孔,MTT加入量为100μg(5mg/ml,20μl);其它实验条件为选用570nm为主波长,630nm为参考波长;96孔板不离心;MTT作用4小时。MTT法经济、准确、快速和简便,从而为临床急性白血病的药物敏感检测奠定方法学基础。
In this experiment, two kinds of in vitro cultured leukemic cell lines (K_(562), HL_(60)) were used to study the MTT experimental conditions. The results showed that there was a linear relationship between the number of metabolites of MTT metabolites. The optimal experimental conditions were as follows: the number of experimental cells was 2×10~5/well or 4×10~5/well, the amount of MTT added was 100 μg (5 mg/ml, 20 μl); the other experimental conditions were the use of 570 nm as the main wavelength. 630nm is the reference wavelength; 96-well plates are not centrifuged; MTT acts for 4 hours. The MTT method is economical, accurate, rapid, and simple, which lays the methodological foundation for the drug sensitive detection of clinical acute leukemia.