Engrafted human stem cell–derived hepatocytes establish an infectious HCV murine model
Arnaud Carpentier, … , Stephen M. Feinstone, T. Jake Liang
Arnaud Carpentier, … , Stephen M. Feinstone, T. Jake Liang
Published October 8, 2014
Citation Information: J Clin Invest. 2014;124(11):4953-4964. https://doi.org/10.1172/JCI75456.
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Technical Advance Hepatology

Engrafted human stem cell–derived hepatocytes establish an infectious HCV murine model

Abstract

The demonstrated ability to differentiate both human embryonic stem cells (hESCs) and patient-derived induced pluripotent stem cells (hiPSCs) into hepatocyte-like cells (HLCs) holds great promise for both regenerative medicine and liver disease research. Here, we determined that, despite an immature phenotype, differentiated HLCs are permissive to hepatitis C virus (HCV) infection and mount an interferon response to HCV infection in vitro. HLCs differentiated from hESCs and hiPSCs could be engrafted in the liver parenchyma of immune-deficient transgenic mice carrying the urokinase-type plasminogen activator gene driven by the major urinary protein promoter. The HLCs were maintained for more than 3 months in the livers of chimeric mice, in which they underwent further maturation and proliferation. These engrafted and expanded human HLCs were permissive to in vivo infection with HCV-positive sera and supported long-term infection of multiple HCV genotypes. Our study demonstrates efficient engraftment and in vivo HCV infection of human stem cell–derived hepatocytes and provides a model to study chronic HCV infection in patient-derived hepatocytes, action of antiviral therapies, and the biology of HCV infection.

Authors

Arnaud Carpentier, Abeba Tesfaye, Virginia Chu, Ila Nimgaonkar, Fang Zhang, Seung Bum Lee, Snorri S. Thorgeirsson, Stephen M. Feinstone, T. Jake Liang

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Figure 6

In situ maturation of engrafted HLCs.

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In situ maturation of engrafted HLCs. (A and B) Coimmunostaining for hAL...
(A and B) Coimmunostaining for hALB, hAAT, and hAFP on liver sections of engrafted mice 100 days pe, visualized by confocal microscopy. Asterisks indicate central veins. Original magnification, ×10; scale bar: 400 μm. Images at ×20 magnification are shown in Supplemental Figure 6. (C) IFA for hALB, hAFP, and hAAT 14 days pe. Scale bar: 200 μm. (D) ELISA for hAFP in sera of engrafted mice during the first 2 weeks pe (black lines) and control nonengrafted mice (red line). (E and F) Coimmunostaining for hALB and human and mouse CYP450 isoforms, confirming expression of mature hepatic markers in engrafted HLCs. Scale bars: 200 μm. Asterisks indicate central veins.