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.
View: Text | PDF
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

×

Figure 3

In vitro HCV infection of HLCs.

Options: View larger image (or click on image) Download as PowerPoint
In vitro HCV infection of HLCs. (A) Quantification of intracellular and ...
(A) Quantification of intracellular and extracellular HCV RNA by RTqPCR, and quantification of HCV core antigen in the supernatant by ELISA, after infection with JFH1-HCVcc. Inhibition of JFH1-HCV replication by 100 mM 2′-C-methylcytidine (2MC) is indicated in red (red arrows indicate when the 2MC was added). (B) Inoculation of Huh7.5.1 with supernatant of JFH1-HCVcc–infected HLCs, followed by immunofluorescence detection of HCV core protein, indicating production of infectious virus. Scale bar: 100 μm. (C and D) HLCs were transduced to express the IPS-NLS-RFP vector. One day later, cells were inoculated with (C) JFH1-HCVcc– or (D) HCV-positive clinical isolates of different genotypes. Two days pi, cells were observed for relocalization events, confirming infection of the cells by HCVcc and authentic HCV particles from patients’ sera. Scale bars: 50 μm. (E) Time-dependent visualization of HDFR, suggesting cell-to-cell transmission of HCV infection. Scale bars: 50 μm. (F) Induction of an antiviral innate immunity in response to JFH1-HCVcc infection of HLCs, as assessed by RTqPCR. *P < 0.05. Data are expressed as mean ± SEM.