The objective of this study was to analyse the response of hepatocytes on various concentrations of 17 beta-oestradiol (17 beta-E) under iron-induced oxidative stress in vitro. Isolated by in situ collagenase perfusion hepatocytes were cultured in DMEM/HAMS-12 (v/v) medium without any additional agents (control), with Fe3+ alone, and with Fe3+. and 0.2%, 0.02%, and 0.002% solution of 17 beta-E (17 beta-EI, 17 beta-EII, and 17 beta-EIII, respectively). After 24, 48, and 72 h, medium malonylodialdehyde (MDA), haptoglobin (Hpt) concentration and proliferative activity were determined. In comparison to control samples, and samples collected at 24 and 72 h, hepatocytes exposition to Fe3+, caused a significant increase in MDA (0.056 +/- 0.011 nM/mL) only after 48 h of incubation. Each of 17 beta-E concentrations resulted in a decrease in MDA in samples obtained after 24 and 48 it In comparison to the first 24 h. Fe3+. alone and together with 17 beta-EI, 17 beta-EII, and 17 beta-EIII caused a significant augmentation of Hpt level in 48 h and 72 h of the experiment. Each of the 17 beta-E concentrations added to the culture medium resulted in inhibition of proliferative activity, especially in the 72 h of cell culture.