USE OF AN AQUEOUS SOLUBLE TETRAZOLIUM FORMAZAN ASSAY TO MEASURE VIABILITY AND PROLIFERATION OF LYMPHOKINE-DEPENDENT CELL-LINES
USE OF AN AQUEOUS SOLUBLE TETRAZOLIUM FORMAZAN ASSAY TO MEASURE VIABILITY AND PROLIFERATION OF LYMPHOKINE-DEPENDENT CELL-LINES
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DOI：
10.1016/0022-1759(93)90092-l
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发表时间：
1993-01-04
影响因子：
2.2
通讯作者：
OWEN, TC
中科院分区：
文献类型：
--
作者：
BUTTKE, TM;MCCUBREY, JA;OWEN, TC
最近已经描述了一种新的四唑鎓化合物MTS（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）2 H-四唑鎓，内盐），其在吩嗪硫酸甲酯（PMS）存在下被活细胞还原以产生可比色测定的甲瓒产物。MTS/PMS优于其它四唑染料（例如，MTT）是还原甲瓒产物的水溶性，其消除了对去污剂增溶或有机溶剂萃取步骤的需要。与XTT/PMS（另一种产生水溶性甲瓒产物的四唑）相比，其优点包括所产生颜色的吸光度范围（515-580 nm，而不是450 nm）、显色速度快以及MTS/PMS试剂溶液的储存稳定性。在本研究中，MTS/PMS用于测定IL-2依赖的HT-2和CTLL-2细胞系以及IL-3依赖的FDC-PI和FL5.12细胞系的存活率和增殖。对于每种细胞系，甲瓒产物的量是时间依赖性的，并且与活细胞的数量成比例。此外，用HT-2和CTLL-2细胞，发现培养物可以同时用MTS/PMS和[H-3]胸苷标记，染料对后者摄取的影响相对较小。在FDC-P1细胞的研究中进一步利用了这一特征，其中MTS/PMS和[H-3]胸苷的共同添加用于区分细胞活力和增殖。
A new tetrazolium compound, MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)2H-tetrazolium, inner salt), has recently been described which in the presence of phenazine methosulfate (PMS) is reduced by living cells to yield a formazan product that can be assayed colorimetrically. An important advantage of MTS/PMS over other tetrazolium dyes (e.g., MTT) is the aqueous solubility of the reduced formazan product which eliminates the need for detergent solubilization or organic solvent extraction steps. Its advantages over XTT/PMS, another tetrazolium which yields a water-soluble formazan product, include the absorbance range of color produced (515-580 nm as opposed to 450 nm), the rapidity of color development, and the storage stability of the MTS/PMS reagent solution. In the present study, MTS/PMS was used to assay viability and proliferation of the IL-2-dependent HT-2 and CTLL-2 cell lines and the IL-3-dependent FDC-PI and FL5.12 cell lines. With each cell line, the amount of formazan product was time-dependent and proportional to the number of viable cells. Furthermore, with both HT-2 and CTLL-2 cells it was found that cultures could be simultaneously labeled with MTS/PMS and [H-3]thymidine, with relatively little effect of the dye on uptake of the latter. This feature was further capitalized upon in studies with FDC-P1 cells, in which the co-addition of MTS/PMS and [H-3]thymidine was used to distinguish between cell viability and proliferation.