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Long-term controlled immortalization of a primate hepatic progenitor cell line after Simian virus 40 T-Antigen gene transfer

Oncogene volume 24, pages 541–551 (2005)Cite this article

Abstract

Hepatoblasts are bipotent progenitors of both hepatocytes and cholangiocytes. The lack of stable in vitro culture systems for such cells makes it necessary to generate liver progenitor cell lines by means of immortalization. In this study, we describe the long-term behaviour of a clone of simian foetal hepatic progenitor cells immortalized by Simian virus 40 (SV40) large T-antigen (T-Ag) flanked by loxP sites. Immortalization was associated with the re-expression of telomerase activity, which decreased at late passages (population doubling 120) after more than a year in culture. This decrease was concomitant to telomere shortening and karyotypic instability. However, the chromosomes carrying the p53 gene remained intact and long-term immortalized progenitor cells maintained contact inhibition and proliferative properties. They also displayed the features of a normal bipotent phenotype. We constructed a retroviral vector expressing an inducible Cre recombinase and transferred it into the immortalized progenitors. Activation of the Cre recombinase by 4-hydroxy-tamoxifen induced SV40 T-Ag excision, leading to the death of cells expressing Cre recombinase. Immortalized progenitors at late passages stopped growing and eventually disappeared after transplantation into the livers of immunocompromised mice. These cells provide a novel model to study hepatic differentiation and carcinogenesis.

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Acknowledgements

We thank Drs D Mahieu and I Dagher for help in animal surgery, L Cam for helpful discussion and Mrs B Savary for assistance in artwork. This research was supported by the EC, QLK3CT 1999-00364, and grants from Inserm ATC Biothérapies, Association Française contre les Myopathies, the Fondation pour la Recherche Médicale, the Etablissement Français des Greffes, the ‘GIS Maladies Rares’ and the Institut Pasteur. JE Allain was supported by Fondation pour la Recherche Médicale.

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Author notes

  1. Frédérique Capron

    Present address: Service d'anatomo-pathologie, Hôpital de la Pitié-Salpétrière, 75013, Paris, France

Authors and Affiliations

  1. INSERM EMI 00-20; University Paris XI, Hôpital Kremlin-Bicêtre, 80 avenue du Général Leclerc, 94276, Kremlin-Bicêtre, France

    Jean-Paul Delgado, Alexandre Parouchev, Jean-Etienne Allain, Frédérique Capron & Anne Weber

  2. Laboratoire de Radiopathologie, DSV/DRR, CEA, 18 route du Panorama, BP6-92265, Fontenay-aux-Roses, France

    Gaëlle Pennarun, Laurent R Gauthier, Bernard Dutrillaux & François D Boussin

  3. Laboratoire d'Instabilité du Génome et Cancer, CNRS, Institut Curie, 26 rue d'Ulm, 75005, Paris, France

    Anne-Marie Dutrillaux & Bernard Dutrillaux

  4. INSERM EPI 01-01, Institut Pasteur, 25 rue du Dr Roux, 75015, Paris, France

    James Di Santo

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  1. Jean-Paul Delgado
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Correspondence to Anne Weber.

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Delgado, JP., Parouchev, A., Allain, JE. et al. Long-term controlled immortalization of a primate hepatic progenitor cell line after Simian virus 40 T-Antigen gene transfer. Oncogene 24, 541–551 (2005). https://doi.org/10.1038/sj.onc.1208089

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