The effects of culture medium and serum components on immunoglobulin (Ig) production by mouse splenocytes were examined. In this study, we showed that culture medium containing charcoal-dextran-treated fetal bovine serum (CDFBS) supported Ig production more efficiently than culture medium containing FBS. In addition, RPMI 1640 medium supported Ig production more efficiently than Dulbecco modified Eagle medium. In addition, an increase in medium IgA production was observed with the increase in both CDFBS and FBS concentrations, whereas an increase in IgM level was observed in the presence of 5% and 10% FBS and 10% CDFBS. Dose-dependent effects of estrogenic compounds on Ig production were also examined. We found that 17beta-estradiol induced increases in IgM and IgE levels, but diethylstilbestrol enhanced only the IgE level. On the other hand, tamoxifen and bisphenol A enhanced medium IgM level at physiological concentrations. Among flavonoids, daidzein enhanced IgM and IgE levels at concentrations above 10 microM, and genistein induced a decrease in IgM level and an increase in IgE level at concentrations above 10 microm. In addition, quercetin and luteolin enhanced medium IgE level at all concentrations tested, whereas IgA, IgG, and IgM levels were not affected. These results suggest that environmental estrogens affect Ig production of mouse Splenocytes in a complex and class-specific manner.