Unknown

HSG or HGF cells were inoculated into 96- well plates at a density of 4 x 103 cells/well in 0.1 ml of MEM or •- MEM supplemented with 10% FBS and cultured at 37C for 2 days. The medium was replaced by serum-free medium 1 hour before the assay. Phenol compound Bis-MMP (Dimethoxy Di-p-Cresol) (10^-5 to 10-1 M) dissolved in DMSO were added to the cells, giving a final concentration of 10^-7 to 10^-3 M. The final DMSO concentration was 1%. After incubation for 2 days, viability was estimated by the MTT method. 25 mkМ of Cell Titer 96 Aqueous One solution of MTT was added to each well. After incubation for 3 hours at 37ЖC, the absorbance at 492 nm was determined with a microplate reader (Biochromatic; Labsystem, Helsinki, Finland). The viability was defined as the ratio (percent) of the absorbance in the experimental well to that of the control well, which contained 1% DMSO (no test compound). A doseresponse curve of viability (%) was plotted to define the concentration of eugenol-related compounds that reduced MTT formazan production. HSG cells were maintained as monolayer cultures at 37ЖC in MEM medium supplemented with 10% FBS, in a humidified 5% CO2 atmosphere