- Research article
In vitro and in vivo characteristics of hepatic oval cells modified with human hepatocyte growth factor
Cellular & Molecular Biology Letters volume 18, pages 507–521 (2013)
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.
epidermal growth factor
hepatocyte growth factor
human hepatocyte growth factor
hepatic oval cells
leukemia inhibitory factor
median survival time
Faris, R.A., Konkin, T. and Halpert, G. Liver stem cells: a potential source of hepatocytes for the treatment of human liver disease. Artif. Organs 25 (2001) 513–521.
Fausto, N. and Campbell, J.S. The role of hepatocytes and oval cells in liver regeneration and repopulation. Mech. Dev. 120 (2003) 117–130.
Zimmermann, A. Regulation of liver regeneration. Nephrol. Dial. Transplant. 19Suppl 4 (2004) iv6–10.
Oertel, M. and Shafritz, D.A. Stem cells, cell transplantation and liver repopulation. Biochim. Biophys. Acta 1782 (2008) 61–74.
Williams, J.M., Oh, S.H., Jorgensen, M., Steiger, N., Darwiche, H., Shupe, T. and Petersen, B.E. The role of the Wnt family of secreted proteins in rat oval “stem” cell-based liver regeneration: Wnt1 drives differentiation. Am. J. Pathol. 176 (2010) 2732–2742.
Matsumoto, K. and Nakamura, T. Emerging multipotent aspects of hepatocyte growth factor. J. Biochem. 119 (1996) 591–600.
Huh, C.G., Factor, V.M., Sanchez, A., Uchida, K., Conner, E.A. and Thorgeirsson, S.S. Hepatocyte growth factor/c-met signaling pathway is required for efficient liver regeneration and repair. Proc. Natl. Acad. Sci. USA 101 (2004) 4477–4482.
Matsumoto, K. and Nakamura, T. Hepatocyte growth factor: molecular structure, roles in liver regeneration, and other biological functions. Crit. Rev. Oncog. 3 (1992) 27–54.
Hasuike, S., Ido, A., Uto, H., Moriuchi, A., Tahara, Y., Numata, M., Nagata, K., Hori, T., Hayashi, K. and Tsubouchi, H. Hepatocyte growth factor accelerates the proliferation of hepatic oval cells and possibly promotes the differentiation in a 2-acetylaminofluorene/partial hepatectomy model in rats. J. Gastroenterol. Hepatol. 20 (2005) 1753–1761.
Shiota, G., Kunisada, T., Oyama, K., Udagawa, A., Nomi, T., Tanaka, K., Tsutsumi, A., Isono, M., Nakamura, T., Hamada, H., Sakatani, T., Sell, S., Sato, K., Ito, H. and Kawasaki, H. In vivo transfer of hepatocyte growth factor gene accelerates proliferation of hepatic oval cells in a 2-acetylaminofluorene/partial hepatectomy model in rats. FEBS. Lett. 470 (2000) 325–330.
Kato, H., Shimomura, T., Murai, R., Gonda, K., Ishii, K., Yoshida, Y., Kanbe, T., Hashiguchi, K., Sakabe, T., Takubo, K., Okano, J., Tsuchiya, H., Hoshikawa, Y., Kurimasa, A. and Shiota, G. Regulation of hepatic oval cell proliferation by adenoviral mediated hepatocyte growth factor gene transfer and signal transduction inhibitors. Hepatogastroenterology 54 (2007) 821–825.
Li, Z., Li, L., Ran, J.H., Zhang, S.N., Liu, J., Li, L.B. and Chen, J. Experimental study on hepatic oval cells to establish proliferation model and isolation, culture and differentiation in vitro in adult rat. China Journal of Modern Medicine 5 (2010) 31–34.
Savatier, P., Lapillonne, H., van Grunsven, L.A., Rudkin, B.B. and Samarut, J. Withdrawal of differentiation inhibitory activity/leukemia inhibitory factor up-regulates D-type cyclins and cyclin-dependent kinase inhibitors in mouse embryonic stem cells. Oncogene 12 (1996) 309–322.
Leary, A.G., Wong, G.G., Clark, S.C., Smith, A.G. and Ogawa, M. Leukemia inhibitory factor differentiation-inhibiting activity/human interleukin for DA cells augments proliferation of human hematopoietic stem cells. Blood 75 (1990) 1960–1964.
Xie, R. and Xu, S.R. Two-cuff technique was applied in the establishment of orthotopic liver transplantation in rats Journal of Jiangsu University (Medicine Edition) 16 (2006) 392–395.
Li, Z., Chen, J., Li, L., Ran, J.H., Li, X.H., Liu, Z.H., Liu, G.J., Gao, Y.C., Zhang, X.L. and Sun, H.D. Human hepatocyte growth factor (hHGF)-modified hepatic oval cells improve liver transplant survival. PLoS One 7 (2012) e44805.
Ma, P.C., Maulik, G., Christensen, J. and Salgia, R. c-Met: structure, functions and potential for therapeutic inhibition. Cancer Metastasis Rev. 22 (2003) 309–325.
Schmidt, C., Bladt, F., Goedecke, S., Brinkmann, V., Zschiesche, W., Sharpe, M., Gherardi, E. and Birchmeier, C. Scatter factor/hepatocyte growth factor is essential for liver development. Nature 373 (1995) 699–702.
Trusolino, L., Bertotti, A. and Comoglio, P.M. MET signalling: principles and functions in development, organ regeneration and cancer. Nat. Rev. Mol. Cell Biol. 11 (2010) 834–848.
Montalvo-Jave, E.E., Escalante-Tattersfield, T., Ortega-Salgado, J.A., Pina, E. and Geller, D.A. Factors in the pathophysiology of the liver ischemiareperfusion injury. J. Surg. Res. 147 (2008) 153–159.
Kitta, K., Day, R.M., Ikeda, T. and Suzuki, Y.J. Hepatocyte growth factor protects cardiac myocytes against oxidative stress-induced apoptosis. Free Radic. Biol. Med. 31 (2001) 902–910.
Zhu Li and Juan Chen contributed equally to this study
About this article
Cite this article
Li, Z., Chen, J., Li, L. et al. In vitro and in vivo characteristics of hepatic oval cells modified with human hepatocyte growth factor. Cell Mol Biol Lett 18, 507–521 (2013). https://doi.org/10.2478/s11658-013-0104-1
- Hepatic oval cells
- Hepatocyte growth factor
- Plasmid transfection
- Liver transplantation
- In vitro
- Liver regeneration