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Biliary organoids, an innovative three\u2010dimensional (3D) in vitro model, have emerged to bridge this gap, closely mimicking tissue structure and function. This review systematically summarizes the construction methods of biliary organoids\u2014matrix\u2010independent methods and matrix\u2010dependent methods, as well as tissue engineering\u2010based strategies, such as bioprinting and microfluidics\u2014and cell sources, including primary tissues, pluripotent stem cells, and tumor\u2010derived cells. It also explores the potential roles of key signaling pathways that drive biliary development and disease in guiding cell differentiation, proliferation, and tissue organization during biliary organoid construction. It explores recent applications in disease modeling and clinical translation, leveraging gene editing, chemical induction, inflammatory stimulation, and co\u2010culture systems. Despite their potential, challenges persist in model stability, long\u2010term culture, and immune microenvironment simulation. Future advances, integrating multi\u2010omics, dynamic culture systems, and emerging bioengineering technologies, promise to enhance physiological relevance. Biliary organoids are poised to transform fundamental research, drug screening, and personalized medicine, accelerating clinical breakthroughs in hepatobiliary disease management.<\/jats:p>","DOI":"10.1002\/adhm.202501776","type":"journal-article","created":{"date-parts":[[2025,7,18]],"date-time":"2025-07-18T06:45:44Z","timestamp":1752821144000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Advances in Biliary Disease Organoid Research: From Model Construction to Clinical Applications"],"prefix":"10.1002","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-9338-3062","authenticated-orcid":false,"given":"Boming","family":"Peng","sequence":"first","affiliation":[{"name":"Department of Hepatobiliary Surgery Haikou Affiliated Hospital of Central South University Xiangya School of\u00a0Medicine  Haikou 570208 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology Haikou Affiliated Hospital of Central South University Xiangya School of\u00a0Medicine  Haikou 570208 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianquan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Hepatobiliary Surgery Haikou Affiliated Hospital of Central South University Xiangya School of\u00a0Medicine  Haikou 570208 China"},{"name":"Haikou Key Laboratory of Clinical Research and Transformation of Digestive Diseases  Haikou 570208 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-4712-2048","authenticated-orcid":false,"given":"Yang","family":"Xiang","sequence":"additional","affiliation":[{"name":"Department of Hepatobiliary Surgery Haikou Affiliated Hospital of Central South University Xiangya School of\u00a0Medicine  Haikou 570208 China"},{"name":"Haikou Key Laboratory of Clinical Research and Transformation of Digestive Diseases  Haikou 570208 China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,7,17]]},"reference":[{"key":"e_1_2_9_1_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41572-021-00300-2"},{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms222212200"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2021.04.005"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1088\/1758-5090\/aaf9fa"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/jev2.12280"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2023.216180"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-019-0168-0"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2016.097"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2020.108453"},{"key":"e_1_2_9_10_1","first-page":"303","author":"Salas\u2010Silva S.","year":"2023","journal-title":"Biomaterials"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13287-024-03877-z"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2022.121647"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioactmat.2024.08.010"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01911-20"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41556-022-00989-7"},{"key":"e_1_2_9_16_1","unstructured":"K. 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