This is a PDF file of a peer-reviewed paper that has been accepted for publication. Although unedited, the content has been subjected to preliminary formatting. Nature is providing this early version of the typeset paper as a service to our authors and readers. The text and figures will undergo copyediting and a proof review before the paper is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers apply.

Organoid technology holds great promise for regenerative medicine but has not yet been applied to humans. We address this challenge using cholangiocyte organoids in the context of cholangiopathies, which represent a key reason for liver transplantation. Using single-cell RNA sequencing, we show that primary human cholangiocytes display transcriptional diversity that is lost in organoid culture. However, cholangiocyte organoids remain plastic and resume their in vivo signatures when transplanted back in the biliary tree. We then utilize a model of cell engraftment in human livers undergoing ex vivo normothermic perfusion to demonstrate that this property allows extrahepatic organoids to repair human intrahepatic ducts after transplantation. Our results provide proof of principle that cholangiocyte organoids can be used to repair human biliary epithelium.

This is a PDF file of a peer-reviewed paper that has been accepted for publication. Although unedited, the content has been subjected to preliminary formatting. Nature is providing this early version of the typeset paper as a service to our authors and readers. The text and figures will undergo copyediting and a proof review before the paper is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers apply.

Biliary disorders are the leading indication for pediatric liver transplantation due to the lack of alternative treatments for repairing or replacing damaged bile ducts. To address this challenge, 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 function from primary extra-or intrahepatic duct cholangiocytes, within 2 weeks of isolation. COs are subsequently seeded on Poly-Glycolic Acid scaffolds or densified collagen constructs for 4 weeks to generate bioengineered tissue retaining biliary characteristics. Therefore, expertise in organoid culture and tissue-engineering are desirable for optimal results. Importantly, COs correspond to mature functional cholangiocytes, differentiating our method from alternative organoid systems propagating adult stem cells. Consequently, COs provide a unique platform for studies in biliary physiology and pathophysiology; while the resulting bioengineered tissue has broad applications for regenerative medicine and cholangiopathies.