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

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.

A Bile Duct‐on‐a‐Chip With Organ‐Level Functions

“…Isolated perfused intrahepatic bile duct units maintain their in vivo organization and have been used effectively to characterize cholangiocyte cilia as mechanical and osmotic sensors, but they do not permit measurement of monolayer permeability, manipulation of the underlying matrix, or incorporation of genetically modified or other specific populations of cholangiocytes . Cholangiocytes in 3D spheroid culture differentiate well, can be cultured in physiological matrices, and demonstrate barrier functions and transport activities . However, spheroids are highly variable in shape and size, and it is difficult to control their positions in a gel (for microscopy), sample lumen content, or treat the cholangiocyte apical surface.…”

Peribiliary Gland Niche Participates in Biliary Tree Regeneration in Mouse and in Human Primary Sclerosing Cholangitis

“…Furthermore, PBG stem cell niche gives rise to a secondary regenerative response, leading to biliary carcinogenesis in human patients with PSC and in experimental genetically induced biliary injury . Generally, the emerging concept is that PBG stem cell niche can be activated in the context of pathologies affecting larger IHBDs and EHBDs, despite the well‐recognized proliferative capability of mature cholangiocytes . Thus, the actual contribution of BTSCs in the bile duct regeneration remains an open question.…”

“…Bile ducts are lined by specialized epithelial cells named cholangiocytes . Cholangiocytes are involved in the modification of bile composition, and their proliferation is responsible for the turnover of the biliary epithelium . However, human diseases affecting the biliary tree (i.e., cholangiopathies) determine an impairment of cholangiocyte proliferative capabilities .…”

From Hepatocyte to Cholangiocyte: The Remarkable Potential of Transdifferentiation to Treat Cholestatic Diseases

“…The field of regenerative medicine holds great promise for future treatment options in cholestatic liver diseases. Recently, a new method for the propagation of human extrahepatic cholangiocyte organoids that self‐organize into bile duct–like tubes was described, setting the stage for bioengineering of the extrahepatic biliary tree . Similarly, the ability for the hepatocyte to transdifferentiate into a functioning intrahepatic biliary tree is noteworthy and has strong implications for future treatment options for ALGS, as well as other cholestatic diseases characterized by intrahepatic biliary injury, such as primary biliary cholangitis, biliary atresia after Kasai portoenterostomy, and primary sclerosing cholangitis.…”