两种检测细胞活性的改良MTT法

Two improved MTT methods for assaying vitality of cells

四甲基偶氮唑盐(MTT)测定法是依靠线粒体琥珀酸脱氢酶催化MTT形成蓝色甲(月替)的多少来检测活细胞数和功能状态的。对影响甲(月替)溶解的助溶剂配方和用量作了实验比较,提出两种配方和合适用量,使此法的敏感性提高了25%,与~3H-TdR掺入法接近,并且相关性好(P<0.001)。改良MTT法重复性好、操作简便、较安全、费用低,可替代~3H-TdR法用于细胞活性的测定。

3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide(MTT) methodis a simple and convenient one for assaying vitality of cells, however the sensitivity is not so satis-factory as that of ~3H- TdR incorporation. After comparatively studying the modified MTT meth-ods reported, two solubilizers for formazane, which were 0.01 mol/L HCl-10% triton-50%isopropanol (TRITON-ISOP) and 0.01mol/L HCl-iospropanol (ISOP), were established andtwo modified MTT methods were proposed. Ⅰ. The overnight assay needs less labor and pro-cessing time. The same volume of TRITON-ISOP was added to the cell culture, then incubated at37℃ for 12-16 hours and read the OD value at the 570nm. The result corresponds to that of^3H-TdR method and the correlativity of the two methods is great significant (P<0.01). Ⅱ. Theimmediate assay using ISOP as solubilizer may show result more quickly. The volume of ISOP is1.5 times to that of the cell culture. The OD value of test can be read within 20-60 minutes afterthoroughly mixing the reagent by mixer. The two improved methods show high sensitivity andgood reproducibility and may replace ~3H-TdR method in assaying vitality of cells.