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Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice

Nature Biotechnology volume 25, pages 903–910 (2007)Cite this article

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Abstract

Mice that could be highly repopulated with human hepatocytes would have many potential uses in drug development and research applications. The best available model of liver humanization, the uroplasminogen-activator transgenic model, has major practical limitations. To provide a broadly useful hepatic xenorepopulation system, we generated severely immunodeficient, fumarylacetoacetate hydrolase (Fah)-deficient mice. After pretreatment with a urokinase-expressing adenovirus, these animals could be highly engrafted (up to 90%) with human hepatocytes from multiple sources, including liver biopsies. Furthermore, human cells could be serially transplanted from primary donors and repopulate the liver for at least four sequential rounds. The expanded cells displayed typical human drug metabolism. This system provides a robust platform to produce high-quality human hepatocytes for tissue culture. It may also be useful for testing the toxicity of drug metabolites and for evaluating pathogens dependent on human liver cells for replication.

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Figure 1: Engraftment and repopulation of Fah−/−/Rag2−/−/Il2rg−/− mice with human hepatocytes.
Figure 2: Histology and immunohistochemistry of chimeric mice.
Figure 3: Human gene and protein expression in chimeric mice.
Figure 4: Serial transplantation of human hepatocytes.
Figure 5: Absence of cell fusion.
Figure 6: Drug metabolism.

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  • 11 March 2008

    In the HTML version of this article initially published, the statement that the authors had no competing financial interests was incorrect. The authors declare competing financial interests and the details have now been posted online. The error has been corrected in the HTML version of the article.

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Acknowledgements

This work was supported by NIH grant RO1-DK48252 and funds from the Juvenile Diabetes Foundation to M.G. and NIH grants DK 92310 and GM 06346 to S.S.

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Authors and Affiliations

  1. Oregon Stem Cell Center, Oregon Health & Science University, Portland, 97239, Oregon, USA

    Hisaya Azuma, Nicole Paulk, Craig Dorrell, Muhsen Al-Dhalimy & Markus Grompe

  2. Department of Pathology, University of Pittsburgh Medical Center, Pittsburgh, 15261, Pennsylvania, USA

    Aarati Ranade, Ewa Ellis & Stephen Strom

  3. Department of Pediatrics, Stanford University, California, 94305, USA

    Mark A Kay

  4. Department of Pathology, Texas Children's Hospital, Baylor College of Medicine, Houston, 77030, Texas, USA

    Milton Finegold

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  1. Hisaya Azuma
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Contributions

H.A., primary experimenter, performed all transplantations and analyzed repopulated mice; N.P., FISH in Supplementary Figure 2; A.R., E.E., drug metabolism and drug metabolism gene expression; C.D., FACS analysis; M.A.-D., mouse breeding and transplantation assistance; S.S., provided human hepatocytes and supervised drug metabolism work; M.A.K., provided urokinase adenovirus; M.F., all tissue histology; M.G., overall project planning and coordination.

Corresponding author

Correspondence to Markus Grompe.

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Competing interests

M.G. owns stock shares in Yecuris, Inc., which has licensed the FRG liver repopulation technology.

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Azuma, H., Paulk, N., Ranade, A. et al. Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice. Nat Biotechnol 25, 903–910 (2007). https://doi.org/10.1038/nbt1326

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  • DOI: https://doi.org/10.1038/nbt1326

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