Human stem cell-derived hepatocyte-like cells (HLCs) offer an attractive platform to study liver biology. Despite their numerous advantages, HLCs lack critical in vivo characteristics, including cell polarity. Here, we report a stem cell differentiation protocol that uses transwell filters to generate columnar polarized HLCs with clearly defined basolateral and apical membranes separated by tight junctions. We show that polarized HLCs secrete cargo directionally: Albumin, urea, and lipoproteins are secreted basolaterally, whereas bile acids are secreted apically. Further, we show that enterically transmitted hepatitis E virus (HEV) progeny particles are secreted basolaterally as quasi-enveloped particles and apically as naked virions, recapitulating essential steps of the natural infectious cycle in vivo. We also provide proof-of-concept that polarized HLCs can be used for pharmacokinetic and drug-drug interaction studies. This novel system provides a powerful tool to study hepatocyte biology, disease mechanisms, genetic variation, and drug metabolism in a more physiologically relevant setting.
Conflict of interest statement
The authors declare no conflict of interest.
Vectorial Release of Hepatitis E Virus in Polarized Human Hepatocytes.J Virol. 2019 Feb 5;93(4):e01207-18. doi: 10.1128/JVI.01207-18. Print 2019 Feb 15. J Virol. 2019. PMID: 30463960 Free PMC article.
Biliary Secretion of Quasi-Enveloped Human Hepatitis A Virus.mBio. 2016 Dec 6;7(6):e01998-16. doi: 10.1128/mBio.01998-16. mBio. 2016. PMID: 27923925 Free PMC article.
Human stem cell-derived hepatocytes as a model for hepatitis B virus infection, spreading and virus-host interactions.J Hepatol. 2017 Mar;66(3):494-503. doi: 10.1016/j.jhep.2016.10.009. Epub 2016 Oct 14. J Hepatol. 2017. PMID: 27746336 Free PMC article.
[Generation of Zone-specific Hepatocyte-like Cells from Human Induced Pluripotent Stem Cells for Accurate Prediction of Drug-induced Hepatotoxicity].Yakugaku Zasshi. 2019;139(12):1509-1512. doi: 10.1248/yakushi.19-00170. Yakugaku Zasshi. 2019. PMID: 31787637 Review. Japanese.
Stem Cell-Derived Culture Models of Hepatitis E Virus Infection.Cold Spring Harb Perspect Med. 2019 Mar 1;9(3):a031799. doi: 10.1101/cshperspect.a031799. Cold Spring Harb Perspect Med. 2019. PMID: 29686039 Free PMC article. Review.