Ample evidence exists regarding the presence of gender difference in organ transplantation, with female livers having superior outcome to male donor organs. A previous in vitro report using UW solution confirmed these clinical observations. As there is now a mounting interest using HTK solution in liver transplantation, we undertook this study to determine if the gender of the donor affects the function of HTK-preserved livers. Rat livers of either sex were harvested and stored for 24 h with 4 °C cold HTK solution. Storage was followed by 2 h reperfusion with 37 °C warm KHB in an isolated liver perfusion system. Apoptosis was morphologically quantified using fluorescence microscopy of bisbenzimide-stained parenchymal tissue. Trypan blue perfusion allowed for assessment of cell membrane damage, indicating both secondary apoptotic and primary necrotic cell death. As indicators of global graft quality, bile flow, oxygen consumption, K+-excretion and enzyme release were determined. Upon 2 h of reperfusion, livers developed apoptotic as well as necrotic cell death. Livers from male donors showed increased vulnerability to hepatic preservation-reperfusion injury as indicated by increased hepatocellular apoptosis and necrosis, which was associated with a more pronounced deterioration of excretory and metabolic parameters. This might be ascribed to the reported higher activity of histidase in male livers, thus promoting the degradation of histidine, the main buffer in HTK solution. Thus, care may be taken when applying HTK solution for preservation of critical livers from male donors.