Journal Article

Reconstruction of the mouse extrahepatic biliary tree using primary human extrahepatic cholangiocyte organoids

Bertero, Alessandro, Zilbauer, Matthias, Berntsen, Natalie Lie, Snijders, Kirsten E, Gelson, William T H, Tysoe, Olivia C, Alexander, Graeme J, Markaki, Athina E, Sawiak, Stephen, Pedersen, Marianne T, Pawlowski, Matthias, Bargehr, Johannes, Karlsen, Tom H, Jensen, Kim B, Yiangou, Loukia, Gómez-Vázquez, María J, Georgakopoulos, Nikitas, Sinha, Sanjay, Madrigal, Pedro, Davies, Susan E, Casey, John, Upponi, Sara S, Zonneveld, Mariëlle C F, Pirmadjid, Negar, de Brito, Miguel Cardoso, Brown, Stephanie E, Sampaziotis, Fotios, Valestrand, Laura, Shu, Wenmiao, Ross, Alexander, Saeb-Parsy, Kourosh, Wynn, Thomas A, Justin, Alexander W, Gieseck, Richard L, Melum, Espen, Ortmann, Daniel, Godfrey, Edmund M, Skeldon, Gregor M, Rimland, Casey A, Materek, Paulina M, Hannan, Nicholas R F, Vallier, Ludovic, Simonic, Ingrid

2017

Overview

Repair of defects in the common bile duct is hampered by a lack of healthy donor tissue. Developing human extrahepatic cholangiocyte organoids and testing them in mouse models may provide a way to overcome this limitation. 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 .

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 13 / 13/106 / 13/107 / 59/57 / 631/61/2035 / 631/61/490 / Animals / Bile / Bile ducts / Bile Ducts, Extrahepatic - cytology / Bile Ducts, Extrahepatic - injuries / Bile Ducts, Extrahepatic - physiology / Biliary atresia / Biliary tract / Biliary Tract - cytology / Biliary Tract - injuries / Biliary Tract - physiology / Biliary tract diseases / Biodegradability / Biodegradation / Bioengineering / Biomedicine / Cancer Research / Care and treatment / Cell Transplantation / Cholestasis / Cystic Fibrosis Transmembrane Conductance Regulator - metabolism / Epithelial Cells - cytology / Epithelial Cells - drug effects / Epithelial Cells - metabolism / Epithelium / Gallbladder / Gallbladder - injuries / Gallbladder - physiology / gamma-Glutamyltransferase - metabolism / House mouse / Human cell culture / Humans / In Vitro Techniques / Infectious Diseases / Ischemia / Keratin-19 - metabolism / Keratin-7 - metabolism / Metabolic Diseases / Mice / Molecular Medicine / Neurosciences / Occlusion / Organoids / Organoids - cytology / Organoids - drug effects / Organoids - metabolism / Organoids - physiology / Physiological aspects / Reconstruction / Reconstructive surgery / Regeneration - physiology / Regenerative medicine / Secretin - pharmacology / Somatostatin - pharmacology / Stricture / Tissue engineering / Tissue Engineering - methods / Tissue Scaffolds / Transplantation / Tubes