Ligation of a branch of the portal vein results in marked regenerative hypertrophy of the parenchyma of the unligated hepatic lobe, and the ligated lobe shows atrophy. We investigated possible role of hepatic stimulator substance (HSS) produced in the hepatic lobes after portal branch ligation on this regenerative hypertrophy. HSS was extracted from the ligated and the unligated hepatic lobes 24 hours after portal branch ligation in rabbits. Each of the extracts was administrated to mice and hepatic DNA synthesis was compared. The liver treated by the HSS of the ligated lobe showed a significant increase of BrdU labeling index and liver weight in comparison with the liver treated by that of the unligated lobe. These results suggested that HSS produced in ligated lobe plays an important role on regenerative hypertrophy of the unligated lobe after portal branch ligation.