Pediatric liver transplantation is often required as a consequence of biliary disorders because of the lack of alternative treatments for repairing or replacing damaged bile ducts. To address the lack of availability of pediatric livers suitable for transplantation, we developed a protocol for generating bioengineered biliary tissue suitable for biliary reconstruction. Our platform allows the derivation of cholangiocyte organoids (COs) expressing key biliary markers and retaining functions of primary extra- or intrahepatic duct cholangiocytes within 2 weeks of isolation. COs are subsequently seeded on polyglycolic acid (PGA) scaffolds or densified collagen constructs for 4 weeks to generate bioengineered tissue retaining biliary characteristics. Expertise in organoid culture and tissue engineering is desirable for optimal results. COs correspond to mature functional cholangiocytes, differentiating our method from alternative organoid systems currently available that propagate adult stem cells. Consequently, COs provide a unique platform for studies in biliary physiology and pathophysiology, and the resulting bioengineered tissue has broad applications for regenerative medicine and cholangiopathies.

由于缺乏修复或更换受损胆管的替代疗法，胆道疾病通常需要小儿肝移植。为了解决缺乏适合移植的儿科肝脏的问题，我们开发了一种用于生成适合胆道重建的生物工程胆道组织的协议。我们的平台允许在分离后的 2 周内衍生表达关键胆道标志物和保留原代肝外或肝内胆管细胞功能的胆管细胞类器官 (CO)。随后将 COs 接种在聚乙醇酸 (PGA) 支架或致密的胶原结构上 4 周，以产生保留胆道特性的生物工程组织。获得最佳结果需要类器官培养和组织工程方面的专业知识。COs 对应于成熟的功能性胆管细胞，将我们的方法与目前可用的可繁殖成体干细胞的替代类器官系统区分开来。因此，COs 为胆道生理学和病理生理学研究提供了一个独特的平台，由此产生的生物工程组织在再生医学和胆管病方面具有广泛的应用。