Abstract
In order to understand the life cycle of hepatitis B virus (HBV) and to develop efficient anti-HBV drugs, a useful in vitro cell culture system which allows HBV infection and recapitulates virus-host interactions is essential; however, pre-existing in vitro HBV infection models are often problematic. Here, we examined the potential of human induced-pluripotent stem (iPS) cell-derived hepatocyte-like cells (iPS-HLCs) as an in vitro HBV infection model. Expression levels of several genes involved in HBV infection, including the sodium taurocholate cotransporting polypeptide (NTCP) gene, were gradually elevated as the differentiation status of human iPS cells proceeded to iPS-HLCs. The mRNA levels of these genes were comparable between primary human hepatocytes (PHHs) and iPS-HLCs. Following inoculation with HBV, we found significant production of HBV proteins and viral RNAs in iPS-HLCs. The three major forms of the HBV genome were detected in iPS-HLCs by Southern blotting analysis. Anti-HBV agents entecavir and Myrcludex-B, which are a nucleoside analogue reverse transcriptase inhibitor and a synthetic pre-S1 peptide, respectively, significantly inhibited HBV infection in iPS-HLCs. These data demonstrate that iPS-HLCs can be used as a promising in vitro HBV infection model.
Highlights
An in vitro Hepatitis B virus (HBV) infection model which mimics HBV infection in the human liver is crucial to clarify the HBV life cycle and for the development of anti-HBV agents
The mRNA levels of asialoglycoprotein receptor-1 (ASGPR1) and sodium-taurocholate cotransporting peptide (NTCP), which were transmembrane proteins involved in HBV infection[26,27], were gradually elevated depending on the differentiation status and comparable to or slightly higher than those in primary human hepatocytes (PHHs)
These results indicate that induced-pluripotent stem (iPS)-hepatocyte-like cells (HLCs) expressed several genes crucial for HBV infection at levels comparable to those in PHHs
Summary
An in vitro HBV infection model which mimics HBV infection in the human liver is crucial to clarify the HBV life cycle and for the development of anti-HBV agents. Human hepatocyte-like cells differentiated from human embryonic stem (ES) cells and induced-pluripotent stem (iPS) cells have gained much attention due to their promise for regenerative medicines, and due to their potential for modeling drug metabolism and pathogen infection in vitro[10,11,12,13,14]. IPS-HLCs expressed HBV infection-related cellular factors at levels similar to PHHs. iPS-HLCs were successfully infected by HBV, leading to production of HBV antigens and HBV-derived RNAs. In this study, we examined the potential of iPS-HLCs as an in vitro infection model of HBV. IPS-HLCs expressed HBV infection-related cellular factors at levels similar to PHHs. iPS-HLCs were successfully infected by HBV, leading to production of HBV antigens and HBV-derived RNAs These data indicate that HBV would be a promising in vitro HBV infection model
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
