Use of MTT colorimetric assay to measure cell activation

Abstract

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.