OBJECTIVE To test the hypothesis that DHEAS production from DHEA occurs in hepatic cells and that this production is augmented by the presence of sex steroids or insulin. METHODS In vitro prospective experiment. METHODS Academic medical center. METHODS Hepatoma G2 cells cultured in media supplemented with [1] DHEA (10(-5) mol/L) only, [2] DHEA (10(-5) mol/L) + T (10(-6) mol/L), [3] DHEA (10(-5) mol/L) + E(2) (10(-6) mol/L), [4] DHEA (10(-5) mol/L) + dihydrotestosterone (10(-6) mol/L), [5] DHEA (10(-5) mol/L) + insulin (10 ng/mL), or [6] DHEA (10(-5) mol/L) + insulin (100 ng/mL). METHODS Levels of DHEAS in the media were measured at 0, 2, 4, 6, 8, 12, 24, 48, and 72 hours after adding treatments at time-point 0. RESULTS Dehydroepiandrosterone sulfate was first detected in the hepatoma G2 cell culture media at 12 hours of incubation. The cumulative production rate of DHEAS increased linearly until 72 hours of incubation. When compared with the effect of treatment with DHEA only, treatment with DHEA plus T, dihydrotestosterone, or E(2) delayed the cumulative DHEAS production; alternatively, the addition of insulin did not alter DHEAS production. CONCLUSIONS These data suggest that although hepatic cells have the ability of converting DHEA to DHEAS, neither sex steroids nor insulin results in the increased hepatic production of DHEAS.