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Use of Thymidine Kinase Recombinant Adenovirus and Ganciclovir Mediated Mouse Liver Preconditioning for Hepatocyte Xenotransplantation

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Book cover Hepatocyte Transplantation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1506))

Abstract

Hepatocyte transplantation is the best approach to maintain and propagate differentiated hepatocytes from different species. Host liver has to be adapted for transplanted hepatocytes productive engraftment and proliferation being required a chronic liver injury to eliminate host hepatocytes and provide a proliferative advantage to the transplanted hepatocytes. Most valuable mouse models for xenograft hepatocyte transplantation are based on genetically modified animals to cause a chronic liver damage and to limit host hepatocyte regeneration potential. We present a methodology that generates a chronic liver damage and can be applied to any host mouse strain and animal species based on the inoculation of a recombinant adenovirus to express herpes simplex thymidine kinase in host hepatocytes sensitizing them to ganciclovir treatment. This causes a prolonged liver damage that allows hepatocyte transplantation and generation of regenerative nodules in recipient mouse liver integrated by transplanted cells and host sinusoidal. Obtained chimeric animals maintain functional chimeric nodules for several weeks, ready to be used in any study.

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References

  1. Elaut G, Henkens T, Papeleu P et al (2006) Molecular mechanisms underlying the dedifferentiation process of isolated hepatocytes and their cultures. Curr Drug Metab 7:629–660

    Article  CAS  PubMed  Google Scholar 

  2. De Bruyn T, Chatterjee S, Fattah S et al (2013) Sandwich-cultured hepatocytes: utility for in vitro exploration of hepatobiliary drug disposition and drug-induced hepatotoxicity. Expert Opin Drug Metab Toxicol 9:589–616

    Article  CAS  PubMed  Google Scholar 

  3. Fraczek J, Bolleyn J, Vanhaecke T et al (2013) Primary hepatocyte cultures for pharmaco-toxicological studies: at the busy crossroad of various anti-dedifferentiation strategies. Arch Toxicol 87:577–610

    Article  CAS  PubMed  Google Scholar 

  4. Gomez-Lechon MJ, Tolosa L, Conde I et al (2014) Competency of different cell models to predict human hepatotoxic drugs. Expert Opin Drug Metab Toxicol 10:1553–1568

    Article  CAS  PubMed  Google Scholar 

  5. Aldabe R, Suarez-Amaran L, Usai C et al (2015) Animal models of chronic hepatitis delta virus infection host-virus immunologic interactions. Pathogens 4:46–65

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Iannacone M, Guidotti LG (2015) Mouse models of hepatitis B virus pathogenesis. Cold Spring Harb Perspect Med 5. 10.1101/cshperspect.a021477

  7. Vercauteren K, de Jong YP, Meuleman P (2015) Animal models for the study of HCV. Curr Opin Virol 13:67–74

    Article  PubMed  PubMed Central  Google Scholar 

  8. Jaiswal S, Sharma A, Shukla M et al (2014) Novel pre-clinical methodologies for pharmacokinetic drug-drug interaction studies: spotlight on “humanized” animal models. Drug Metab Rev 46:475–493

    Article  CAS  PubMed  Google Scholar 

  9. Scheer N, Wilson ID (2016) A comparison between genetically humanized and chimeric liver humanized mouse models for studies in drug metabolism and toxicity. Drug Discov Today 21:250–63

    Google Scholar 

  10. Kosaka K, Hiraga N, Imamura M et al (2013) A novel TK-NOG based humanized mouse model for the study of HBV and HCV infections. Biochem Biophys Res Commun 441:230–235

    Article  CAS  PubMed  Google Scholar 

  11. Grompe M, Strom S (2013) Mice with human livers. Gastroenterology 145:1209–1214

    Article  PubMed  Google Scholar 

  12. Douglas DN, Kneteman NM (2015) Generation of improved mouse models for the study of hepatitis C virus. Eur J Pharmacol 759:313–325

    Article  CAS  PubMed  Google Scholar 

  13. Sangro B, Mazzolini G, Ruiz M et al (2010) A phase I clinical trial of thymidine kinase-based gene therapy in advanced hepatocellular carcinoma. Cancer Gene Ther 17:837–843

    Article  CAS  PubMed  Google Scholar 

  14. Moreno D, Balasiddaiah A, Lamas O et al (2013) Usage of adenovirus expressing thymidine kinase mediated hepatocellular damage for enabling mouse liver repopulation with allogenic or xenogenic hepatocytes. PLoS One 8:e74948

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Balasiddaiah A, Moreno D, Guembe L et al (2013) Hepatic differentiation of mouse iPS cells and analysis of liver engraftment potential of multistage iPS progeny. J Physiol Biochem 69:835–845

    Article  CAS  PubMed  Google Scholar 

  16. Duret C, Moreno D, Anangi B et al (2015) Cold-preservation of human adult hepatocytes for liver cell therapy. Cell Transplant. [Epub Ahead of Print]

    Google Scholar 

  17. Caselmann WH (1994) HBV and HDV replication in experimental models: effect of interferon. Antiviral Res 24:121–129

    Article  CAS  PubMed  Google Scholar 

  18. Martin PL, Weinbauer GF (2010) Developmental toxicity testing of biopharmaceuticals in nonhuman primates: previous experience and future directions. Int J Toxicol 29:552–568

    Article  CAS  PubMed  Google Scholar 

  19. Crettaz J, Otano I, Ochoa-Callejero L et al (2009) Treatment of chronic viral hepatitis in woodchucks by prolonged intrahepatic expression of interleukin-12. J Virol 83:2663–2674

    Article  CAS  PubMed  Google Scholar 

  20. van Rijn RS, Simonetti ER, Hagenbeek A et al (2003) A new xenograft model for graft-versus-host disease by intravenous transfer of human peripheral blood mononuclear cells in RAG2-/- gammac-/- double-mutant mice. Blood 102:2522–2531

    Article  CAS  PubMed  Google Scholar 

  21. Viswanathan P, Kapoor S, Kumaran V et al (2014) Etanercept blocks inflammatory responses orchestrated by TNF-alpha to promote transplanted cell engraftment and proliferation in rat liver. Hepatology 60:1378–1388

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Correspondence to Rafael Aldabe .

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Moreno, D., Neri, L., Vicente, E., Vales, A., Aldabe, R. (2017). Use of Thymidine Kinase Recombinant Adenovirus and Ganciclovir Mediated Mouse Liver Preconditioning for Hepatocyte Xenotransplantation. In: Stock, P., Christ, B. (eds) Hepatocyte Transplantation. Methods in Molecular Biology, vol 1506. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6506-9_12

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  • DOI: https://doi.org/10.1007/978-1-4939-6506-9_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6504-5

  • Online ISBN: 978-1-4939-6506-9

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