citations

                    An hepatitis B and D virus infection model using human pluripotent stem cell-derived hepatocytes
                

Authors:

Huanting Chi, Bingqian Qu, Angga Prawira, Talisa Richardt, Lars Maurer, Jungen Hu, Rebecca M Fu , Florian A Lempp, Zhenfeng Zhang , Dirk Grimm, Xianfang Wu, Stephan Urban, Viet Loan Dao Thi

In:

Source: EMBO Rep
Publication Date: (2024)
Issue: 25: 10

Research Area:

Gastroenterology

Stem Cells

Cells used in publication:

Induced Pluripotent Stem Cell (iPS), human: Species: human; Tissue Origin:

Culture Media:

Hepatocyte Culture Medium

Experiment

Generation of human pluripotent stem cell-derived hepatocyte-like cells (HLCs)

The hESC line WA09 (WiCell) was cultured in mTeSR1 medium (STEMCELL Technologies) on Matrigel (Corning) coated plates. WA09 cells were differentiated to definitive endoderm (DE) using the STEMdiff^TM Definitive Endoderm Kit (STEMCELL Technologies) according to the manufacturer’s protocol. To induce hepatic differentiation, DE cells were differentiated in the basal medium consisting of CTS^TM KnockOut^TM DMEM/F-12 (Gibco), 10% KnockOut Serum Replacement (KOSR, Gibco), 1% MEM solution of non-essential amino acids (NEAA, Gibco), 1% GlutaMAX supplement (Gibco), and 1% penicillin–streptomycin (Gibco), supplemented with human hepatocyte growth factor (HGF, Prepotech), Dimethylsulfoxide (DMSO, Sigma-Aldrich), and dexamethasone (Sigma) as previously described (Dao Thi et al, 2020). For final maturation, HLCs were cultured in the Hepatocyte Culture Medium BulletKit^TM (HCM, Lonza) supplemented with 20?ng/mL of recombinant human oncostatin M (OSM, R&D systems). For the co-culture experiment, WA09 cells were transduced with lentivirus to express ZsGreen and selected with 1?µg/mL puromycin as described (Dao Thi et al, 2016) prior to HLC differentiation. prior to HLC differentiation.

Abstract

Current culture systems available for studying hepatitis D virus (HDV) are suboptimal. In this study, we demonstrate that hepatocyte-like cells (HLCs) derived from human pluripotent stem cells (hPSCs) are fully permissive to HDV infection across various tested genotypes. When co-infected with the helper hepatitis B virus (HBV) or transduced to express the HBV envelope protein HBsAg, HLCs effectively release infectious progeny virions. We also show that HBsAg-expressing HLCs support the extracellular spread of HDV, thus providing a valuable platform for testing available anti-HDV regimens. By challenging the cells along the differentiation with HDV infection, we have identified CD63 as a potential HDV co-entry factor that was rate-limiting for HDV infection in immature hepatocytes. Given their renewable source and the potential to derive hPSCs from individual patients, we propose HLCs as a promising model for investigating HDV biology. Our findings offer new insights into HDV infection and expand the repertoire of research tools available for the development of therapeutic interventions.

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