Tran Bang M, Earnest Linda, Flanagan Dustin J, Moselen Jean M, Tran Hoanh, Torresi Joseph, Vincan Elizabeth
Department of Infectious Diseases, University of Melbourne at the Doherty Institute for Infection and Immunity, Melbourne, VIC, Australia.
Department of Microbiology and Immunology, University of Melbourne at the Doherty Institute for Infection and Immunity, Melbourne, VIC, Australia.
Methods Mol Biol. 2025;2951:221-244. doi: 10.1007/7651_2025_626.
The hepatitis B virus (HBV) only robustly infects primary human hepatocytes. This strict viral host and cell tropism has hampered the development of physiologically relevant in vitro culture models of HBV infection. Primary human hepatocytes (PHH) are robustly infected by HBV but are short-lived in tissue culture and rapidly lose their hepatocyte characteristics. Human tissue-derived liver organoids are a novel in vitro physiologically relevant model that supports infection by HBV and mitigates the limitations of PHH. Liver organoids are established by placing tissue fragments into a three-dimensional (3D) basement membrane-rich matrix dome bathed in medium containing supplements and growth factors to support organoid growth. The organoids can be expanded in vitro, cryopreserved, and are genetically stable. The expansion phase organoids, once differentiated to a hepatocyte phenotype, support HBV infection. We couple liver organoids with an adenoviral delivery system to achieve robust HBV infection. This robust model supports the full HBV virus replication cycle.
乙型肝炎病毒（HBV）仅能有效地感染原代人肝细胞。这种严格的病毒宿主和细胞嗜性阻碍了与HBV感染相关的生理相关体外培养模型的发展。原代人肝细胞（PHH）能被HBV有效感染，但在组织培养中寿命较短，并迅速丧失其肝细胞特征。人组织来源的肝类器官是一种新型的体外生理相关模型，可支持HBV感染并减轻PHH的局限性。肝类器官是通过将组织碎片置于富含三维（3D）基底膜的基质穹顶中建立的，该基质穹顶浸浴在含有补充剂和生长因子的培养基中以支持类器官生长。类器官可以在体外扩增、冷冻保存，并且基因稳定。一旦分化为肝细胞表型，扩增期类器官就能支持HBV感染。我们将肝类器官与腺病毒递送系统相结合，以实现有效的HBV感染。这种有效的模型支持完整的HBV病毒复制周期。
