Gerlier D, Thomasset N
J Immunol Methods. 1986 Nov 20;94(1-2):57-63. doi: 10.1016/0022-1759(86)90215-2.
The MTT tetrazolium salt colorimetric assay previously described by Mosmann (1983, J. Immunol. Methods 65, 55) to measure cytotoxicity and cell proliferation was further explored to extend its application to the measurement of cell activation. The level of MTT cleavage by viable cells of various origins was found to be directly proportional to the number of cells but to increase as a non-linear function of time. This non-linear relationship was related to a time-linear cell death during MTT incubation. The cleavage of MTT by viable cells was found to follow first order kinetics and could be fitted to Michaelis' kinetics. Different cell types exhibited similar apparent Km values (1949 microM) and different apparent maximal velocities (V). The apparent V values determined for a given cell type under different experimental conditions were rigorously similar. This analysis of MTT cleavage by viable cells suggests that the colorimetric MTT test can be useful to quantify the activation level of cells, independently of proliferation.
先前由莫斯曼（1983年，《免疫学方法杂志》65卷，55页）描述的用于测量细胞毒性和细胞增殖的MTT四唑盐比色法，被进一步探索以扩展其在细胞活化测量中的应用。发现各种来源的活细胞对MTT的裂解水平与细胞数量成正比，但随着时间的非线性函数增加。这种非线性关系与MTT孵育期间的时间线性细胞死亡有关。发现活细胞对MTT的裂解遵循一级动力学，并且可以拟合米氏动力学。不同细胞类型表现出相似的表观Km值（1949微摩尔）和不同的表观最大速度（V）。在不同实验条件下为给定细胞类型测定的表观V值严格相似。对活细胞MTT裂解的这种分析表明，比色MTT试验可用于独立于增殖来量化细胞的活化水平。
