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  • Viral Transfer Technology
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A preclinical model of hepatocyte gene transfer: the in vivo, in situ perfused rat liver

Gene Therapy volume 7, pages 1816–1823 (2000)Cite this article

Abstract

Delivering retroviruses targeted to hepatocytes in vivo involves the injection of retroviruses directly into the portal vein. The aim of this work was to establish a clinically relevant system for retrovirus-mediated gene transfer in a new model of in vivo, in situ perfused rat liver and to study the transgene expression. At 24 h after partial hepatectomy, the liver was completely excluded from the splanchnic circulation using an extracorporeal shunt. Two independent normothermal, oxygenated perfusion systems were used. First, liver perfusion was carried out with a recirculating system (1 h). Culture supernatant containing retroviruses (1.5 × 108 ffu/ml, β-galactosidase gene) was used as perfusate. Then the liver perfusion was maintained for more 30 min in a single liver passage system using culture medium without retroviruses as perfusate. High hepatocyte transduction rates (up to 34.4%) were obtained. PCR analysis showed no provirus in extrahepatic organs. Viral titrations performed simultaneously (inflow and outflow liver lines) showed that after 1 h of perfusion (up to 30 successive liver passages) retroviruses were still detected in the liver outflow perfusate (up to 2.0 × 107 ffu/ml). Washing the liver for 30 min dramatically decreased the leakage of retroviruses in the outflow. In order to be of clinical use, the injection of retroviruses targeted to hepatocytes in vivo should be done while the liver is completely excluded from the splanchnic circulation to avoid any extrahepatic retrovirus diffusion.

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Acknowledgements

We are grateful to Dr Andrea Pavirani for kindly providing packaging cell line. We thank Dr Horst Homann, Nathalie Ba and Yves Poitevin for their expert technical assistance.

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Author notes

  1. J L De Godoy

    Present address: Disciplina de Cirurgia Pediátrica, Hospital de Clínicas, Universidade Federal do Paraná, Curitiba, Brazil

  2. R Malafosse and M Fabre: RM and MF contributed equally to this work

Authors and Affiliations

  1. Laboratoire de Recherche Chirurgicale et de Biologie Hépatiques, CRI INSERM 94–05, Faculté de Médecine Cochin-Port Royal, Université Paris V, Paris, France

    J L De Godoy, R Malafosse, D Houssin & O Soubrane

  2. Laboratoire d'Anatomie-Pathologique, CHU de Bicêtre, France

    M Fabre

  3. INSERM U 129, Paris, France

    C Mitchell

  4. Transgène, Strasbourg, France

    M Mehtali

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  1. J L De Godoy
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De Godoy, J., Malafosse, R., Fabre, M. et al. A preclinical model of hepatocyte gene transfer: the in vivo, in situ perfused rat liver. Gene Ther 7, 1816–1823 (2000). https://doi.org/10.1038/sj.gt.3301313

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  • DOI: https://doi.org/10.1038/sj.gt.3301313

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