A Robust Human Liver Organoid Model of Hepatitis B Virus Infection.

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2025-04-15 DOI:10.1007/7651_2025_626

Bang M Tran, Linda Earnest, Dustin J Flanagan, Jean M Moselen, Hoanh Tran, Joseph Torresi, Elizabeth Vincan

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引用次数: 0

Abstract

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.

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乙型肝炎病毒感染的人类肝类器官模型。

乙型肝炎病毒（HBV）仅强烈感染原代人肝细胞。这种严格的病毒宿主和细胞趋向性阻碍了HBV感染生理相关体外培养模型的发展。原代人肝细胞（PHH）被HBV强烈感染，但在组织培养中寿命很短，并迅速失去其肝细胞特征。人组织来源的肝类器官是一种新的体外生理相关模型，支持HBV感染并减轻PHH的局限性。肝类器官的建立是通过将组织碎片放入富含基底膜的三维基质穹顶中，并浸泡在含有补充剂和生长因子的培养基中，以支持类器官的生长。类器官可以在体外扩增，冷冻保存，并且具有遗传稳定性。扩张期的类器官，一旦分化为肝细胞表型，支持HBV感染。我们将肝类器官与腺病毒传递系统相结合，以实现强大的HBV感染。这个强大的模型支持整个HBV病毒复制周期。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.