Wellcome Trust-Medical Research Council Stem Cell Institute, Cambridge Stem Cell Institute, Anne McLaren Laboratory, University of Cambridge, Cambridge, UK.
Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, UK.
Nat Med. 2017 Aug;23(8):954-963. doi: 10.1038/nm.4360. Epub 2017 Jul 3.
The treatment of common bile duct (CBD) disorders, such as biliary atresia or ischemic strictures, is restricted by the lack of biliary tissue from healthy donors suitable for surgical reconstruction. Here we report a new method for the isolation and propagation of human cholangiocytes from the extrahepatic biliary tree in the form of extrahepatic cholangiocyte organoids (ECOs) for regenerative medicine applications. The resulting ECOs closely resemble primary cholangiocytes in terms of their transcriptomic profile and functional properties. We explore the regenerative potential of these organoids in vivo and demonstrate that ECOs self-organize into bile duct-like tubes expressing biliary markers following transplantation under the kidney capsule of immunocompromised mice. In addition, when seeded on biodegradable scaffolds, ECOs form tissue-like structures retaining biliary characteristics. The resulting bioengineered tissue can reconstruct the gallbladder wall and repair the biliary epithelium following transplantation into a mouse model of injury. Furthermore, bioengineered artificial ducts can replace the native CBD, with no evidence of cholestasis or occlusion of the lumen. In conclusion, ECOs can successfully reconstruct the biliary tree, providing proof of principle for organ regeneration using human primary cholangiocytes expanded in vitro.
治疗胆总管（CBD）疾病，如胆道闭锁或缺血性狭窄，受到缺乏适合手术重建的健康供体胆管组织的限制。在这里，我们报告了一种从肝外胆管中分离和培养人胆管细胞的新方法，以形成用于再生医学应用的肝外胆管细胞类器官（ECOs）。所得到的 ECOs 在转录组图谱和功能特性方面与原代胆管细胞非常相似。我们探索了这些类器官在体内的再生潜力，并证明 ECOs 在免疫缺陷小鼠的肾囊下移植后可以自组织成表达胆管标记物的胆管样管。此外，当接种在可生物降解的支架上时，ECOs 形成保留胆管特征的组织样结构。所得到的生物工程组织可以在损伤的小鼠模型中移植后重建胆囊壁并修复胆管上皮。此外，生物工程人工胆管可以替代天然 CBD，没有胆管阻塞或管腔阻塞的证据。总之，ECOs 可以成功地重建胆管系统，为使用体外扩增的人原代胆管细胞进行器官再生提供了原理证明。
