Applications of MTT assay to primary cultured rat hepatocytes.

We investigated the optimal conditions for the application of the MTT assay to determine the viability of cell in primary cultures of rat hepatocytes. The optimal conditions were found to be: inoculation at cell density of 1.31 x 10(5) cells/cm2; concentration of 3-(4,5-dimethylthiazoyl-2-yl)-2,5-diphenyltetrazolium bromide (MTT) 0.12 mM; and MTT incorporation time, 2 h or more. We also found that insulin and glucocorticoid, which are usually added to the medium to maintain cell function, had no effect on the viability of the primary cultures of rat hepatocytes.