Abstract
Hepatocyte growth factor (HGF) is a multifunctional growth factor that controls cell scattering. It has been suggested that it regulates the proliferation of hepatic oval cells (HOCs). Using a HOC line that stably expresses the human HGF gene (hHGF), we investigated the in vitro proliferation and differentiation characteristics of hHGF-modified HOCs and explored their potential capacity for intrahepatic transplantation. A modified 2-acetylaminofluorene and partial hepatectomy (2-AAF/PH) model was established to activate the proliferation of oval cells in the rat liver. HOCs were transfected with the pBLAST2-hHGF plasmid and hHGF-carrying HOCs were selected based on blasticidin resistance. The level of hHGF secretion was determined via ELISA. Cell proliferation was determined using the MTT assay. Differentiation was induced by growth factor withdrawal. A two-cuff technique was used for orthotopic liver transplantation, and HOCs or hHGF-modified HOCs were transplanted into the recipients. The levels of biochemical indicators of liver function were measured after transplantation. An HOC line stably expressing hHGF was established. The transfected line showed greater hHGF secretion than normal HOCs. The hHGF gene promoted the proliferation capability of HOCs by reducing the peak time in vitro. The hHGF-modified HOCs differentiated into hepatocytes and bile duct epithelial cells upon growth factor withdrawal in vitro. In addition, hHGF-modified HOC transplantation significantly prolonged the median survival time (MST) and improved the liver function of recipients compared to HOC transplant recipients and nontransplanted controls. Our results indicate that hHGF-modified HOCs may have valuable properties for therapeutic liver regeneration after orthotopic liver transplantation.
Key words: Hepatic oval cells, Hepatocyte growth factor, pBLAST2-hHGF, Plasmid transfection, Proliferation, Differentiation, Liver transplantation, Rats, In vitro, Liver regeneration
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Abbreviations used
- 2-AAF
2-acetylaminofluorene
- ALB
albumin
- ALP
alkaline phosphate
- ALT
alanine aminotransferase
- ChE
cholinesterase
- DBil
direct bilirubin
- EGF
epidermal growth factor
- ELISA
enzyme-linked immunoassay
- GGT
γ-glutamyltransferase
- HGF
hepatocyte growth factor
- hHGF
human hepatocyte growth factor
- HOCs
hepatic oval cells
- I/R
ischemia/reperfusion
- LIF
leukemia inhibitory factor
- MST
median survival time
- PH
partial hepatectomy
Footnotes
Zhu Li and Juan Chen contributed equally to this study
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