Advances in Biliary Disease Organoid Research: From Model Construction to Clinical Applications.

The biliary system is vital to hepatobiliary function, yet diseases like primary sclerosing cholangitis, biliary atresia, cystic fibrosis, and cholangiocarcinoma remain poorly understood due to the limitations of traditional two-dimensional (2D) cell cultures and animal models, which fail to replicate complex human biliary physiology. Biliary organoids, an innovative three-dimensional (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-matrix-independent methods and matrix-dependent methods, as well as tissue engineering-based strategies, such as bioprinting and microfluidics-and cell sources, including primary tissues, pluripotent stem cells, and tumor-derived 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-culture systems. Despite their potential, challenges persist in model stability, long-term culture, and immune microenvironment simulation. Future advances, integrating multi-omics, 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.