1. Reconstruction of the mouse extrahepatic biliary tree using primary human extrahepatic cholangiocyte organoids.
- Author
-
Sampaziotis F, Justin AW, Tysoe OC, Sawiak S, Godfrey EM, Upponi SS, Gieseck RL 3rd, de Brito MC, Berntsen NL, Gómez-Vázquez MJ, Ortmann D, Yiangou L, Ross A, Bargehr J, Bertero A, Zonneveld MCF, Pedersen MT, Pawlowski M, Valestrand L, Madrigal P, Georgakopoulos N, Pirmadjid N, Skeldon GM, Casey J, Shu W, Materek PM, Snijders KE, Brown SE, Rimland CA, Simonic I, Davies SE, Jensen KB, Zilbauer M, Gelson WTH, Alexander GJ, Sinha S, Hannan NRF, Wynn TA, Karlsen TH, Melum E, Markaki AE, Saeb-Parsy K, and Vallier L
- Subjects
- Animals, Bile Ducts, Extrahepatic cytology, Bile Ducts, Extrahepatic injuries, Biliary Tract cytology, Biliary Tract injuries, Biliary Tract physiology, Cell Transplantation, Cystic Fibrosis Transmembrane Conductance Regulator metabolism, Epithelial Cells drug effects, Epithelial Cells metabolism, Gallbladder injuries, Humans, In Vitro Techniques, Keratin-19 metabolism, Keratin-7 metabolism, Mice, Organoids cytology, Organoids drug effects, Organoids metabolism, Secretin pharmacology, Somatostatin pharmacology, Tissue Scaffolds, gamma-Glutamyltransferase metabolism, Bile Ducts, Extrahepatic physiology, Epithelial Cells cytology, Gallbladder physiology, Organoids physiology, Regeneration physiology, Tissue Engineering methods
- Abstract
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.
- Published
- 2017
- Full Text
- View/download PDF