Background/Aims: Although hepatoma-derived growth factor (HDGF) is known to be up-regulated in hepatectomized livers, the regulatory role of HDGF in liver regeneration remains unclear. This study aimed to elucidate the possible role of HDGF in remodeling parenchymal cell growth, microvasculatures, and biliary tree during liver regeneration. Methods: Wild-type (WT) C57BL/6 and HDGF gene knockout (KO) mice received 70% partial hepatectomy (PHx) surgery. Liver plasma and tissues collected after 0, 6h, 1d, 3d, and 7d were used for clincopathological biochemistry, histological, and molecular analyses, including ELISA, qPCR, and Western blotting. Results: Despite no difference in restoration of liver mass between WT and KO mice, plasma levels of AST, ALT, and total bilirubin in KO mice were all significantly higher than those in WT counterparts. KO mice expressed lower mRNA levels of hepatocyte growth factor (HGF), TGF-β1, VEGF, VEGF-R1, and VEGF-R2 in both normal and regenerating livers and had lower HGF in plasma compared to WT controls. HDGF-KO livers at 7 days after PHx had significantly lower protein abundance of CD31, EGFR, NOTCH1 compared to WT controls, suggesting that HDGF gene depletion hampered liver regeneration through interventions in HGF, VEGF, EGFR pathways. Conclusion: HDGF up-regulation in regenerative livers may contribute to the remodeling of hepatic microvasculature and biliary tree, in addition to the repopulation of parenchymal hepatocytes.