USE OF AN AQUEOUS SOLUBLE TETRAZOLIUM FORMAZAN ASSAY FOR CELL-GROWTH ASSAYS IN CULTURE
USE OF AN AQUEOUS SOLUBLE TETRAZOLIUM FORMAZAN ASSAY FOR CELL-GROWTH ASSAYS IN CULTURE
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DOI：
10.3727/095535491820873191
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发表时间：
1991-01-01
影响因子：
16.2
通讯作者：
CORY, JG
中科院分区：
文献类型：
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作者：
CORY, AH;OWEN, TC;CORY, JG
一种新的四唑类似物3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑（MTT）作为MTT的替代品在体外细胞生长的微量培养筛选试验中进行了评价。这种新的四唑化合物，3-（4，5-二甲基噻唑-2-基）-5-（3-羧基甲氧基苯基）-2-（4-磺基苯基）-2H-四唑内盐（MTS），在吩嗪硫酸甲酯（PMS）存在下，在磷酸盐缓冲盐水中产生在490-500 nm处具有最大吸收的水溶性甲瓒产物。形成的有色产物的量与细胞数量和细胞与MTS/PMS孵育的时间成比例。MTS/PMS对小鼠白血病L1210细胞、小鼠埃利希瘤细胞均呈阳性反应。小鼠3 T3成纤维细胞和人结肠肿瘤细胞（HT-29）。HT-29和3 T3成纤维细胞减少MTS/PMS比减少MTT更有效。与由MTT形成的产物的量相比，MTS/PMS给出优异的产物形成。使用MTS/PMS获得的吡唑并咪唑的IC 50值为200 μ M; 5-氟-2 '-脱氧尿苷的IC 50值为200 μ M。IC 50值为0.9 nM。这些值与通过直接细胞计数获得的IC 50值相比非常有利。此外，当在不同孵育时间后测定96孔板中甲瓒产物的半衰期时，获得相同的IC 50值。
A new tetrazolium analog of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was evaluated as a substitute for MTT in the microculture screening assay for in vitro cell growth. This new tetrazolium compound, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS), in the presence of phenazine methosulfate (PMS), gave a water-soluble formazan product that had an absorbance maximum at 490-500 nm in phosphate-buffered saline. The amount of colored product formed was proportional to the number of cells and the time of incubation of the cells with MTS/PMS. MTS/PMS was reactive in all the cell lines tested which included mouse leukemia L1210 cells, mouse Ehrlich tumor cells. mouse 3T3 fibroblasts, and human colon tumor cells (HT-29). HT-29 and 3T3 fibroblasts reduced MTS/PMS more efficiently than they reduced MTT. Comparable to the amount of product formed from MTT, MTS/PMS gave excellent product formation. The IC50 value for pyrazoloimidazole obtained using MTS/PMS was 200-mu-M; for 5-fluoro-2'-deoxyuridine. the IC50 value was 0.9 nM. These values compared very favorably with the IC50 values obtained by direct cell counts. Further, the same IC50 values were obtained when the absorbances of the formazan product in the 96-well plates were determined after different times of incubation.