3D microfluidic liver cultures as a physiological preclinical tool for hepatitis B virus infection

Abstract

With more than 240 million people infected, hepatitis B virus (HBV) is a major health concern. The inability to mimic the complexity of the liver using cell lines and regular primary human hepatocyte (PHH) cultures pose significant limitations for studying host/pathogen interactions. Here, we describe a 3D microfluidic PHH system permissive to HBV infection, which can be maintained for at least 40 days. This system enables the recapitulation of all steps of the HBV life cycle, including the replication of patient-derived HBV and the maintenance of HBV cccDNA. We show that innate immune and cytokine responses following infection with HBV mimic those observed in HBV-infected patients, thus allowing the dissection of pathways important for immune evasion and validation of biomarkers. Additionally, we demonstrate that the co-culture of PHH with other non-parenchymal cells enables the identification of the cellular origin of immune effectors, thus providing a valuable preclinical platform for HBV research.

Decellularized human liver scaffold‐based three‐dimensional culture system facilitate hepatitis B virus infection

“…20 Another group applied the self-assembling (SACC) PHH culture system for HBV persistence infection and preclinical studies. PHH is the best cell model for HBV studies.…”

3D Primary Hepatocyte Culture Systems for Analyses of Liver Diseases, Drug Metabolism, and Toxicity: Emerging Culture Paradigms and Applications

“…Cells in this model remain viable and functional for at least seven days, as judged by calcein acetoxymethyl (AM) and albumin staining, respectively. The model has been used for a variety of applications, including pharmacokinetic studies, evaluation of hepatotoxicity, modeling of nonalcoholic fatty liver disease (NAFLD), and hepatitis B viral infections . In addition, a multitude of small companies and start‐ups provide liver‐on‐a‐chip platforms for which at present very limited peer‐reviewed benchmarking data are available.…”

“…B) PHH cultured as liver‐on‐chip are susceptible to hepatitis B virus (HBV) infections as judged by immunofluorescence of HBV viral antigens (HBsAg and HBcAg) ten days after infection. Reproduced with permission . Copyright 2018, Nature Publishing Group.…”

“…HBV infection has been difficult to model in human liver cell lines as most of these lack the necessary factors for HBV entry. Ortega‐Prieto et al recently established a promising 3D microfluidic system where PHH were permissive to patient‐derived HBV at low titers. Cocultivation with KCs allowed the identification of cell‐specific immune responses and opened the possibility to compare host factors for HBV susceptibility.…”

Insights for clinical diagnostic indicators of virus and host in chronic hepatitis B infection

“…The findings in liver tissues of chimpanzees and CHB patients as well as experiments in vitro have shown that the endogenous IFNα and interferon‐stimulated genes (ISGs) were all not induced while HBV infection . Some research reported that the level of IFNα released by HBV‐infected PHH cells was even decreased and the expression of antiviral genes was hardly observed even in co‐culture with Kupffer cells (KC) …”