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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)Cite this article

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.

Abbreviations

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

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Authors and Affiliations

  1. Department of Hepatobiliary Surgery, Liaocheng People’s Hospital, No. 67 Dongchang West Road, Liaocheng, 252000, Shandong, China

    Zhu Li, Juan Chen, Xue-Hua Li, Zhi-Heng Liu, Gui-Jie Liu, Yan-Chao Gao, Xue-Li Zhang & Hiu-Dong Sun

  2. The First People’s Hospital of Kunming, No. 504 Qinnian Road, Kunming, 650011, Yunnan, China

    Li Li & Jiang-Hua Ran

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  1. Zhu Li
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  2. Juan Chen
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  3. Li Li
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  10. Hiu-Dong Sun
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Correspondence to Zhu Li.

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Zhu Li and Juan Chen contributed equally to this study

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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

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  • DOI: https://doi.org/10.2478/s11658-013-0104-1

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Cellular & Molecular Biology Letters

ISSN: 1689-1392

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