Directing Cholangiocyte Morphogenesis in Natural Biomaterial Scaffolds

Patients with Alagille syndrome carry monogenic mutations in the Notch signaling pathway and face complications such as jaundice and cholestasis. Given the presence of intrahepatic ductopenia in these patients, Notch2 receptor signaling has been implicated in driving normal biliary development and downstream branching morphogenesis. As a result,in vitromodel systems of liver epithelium are needed to further mechanistic insight of biliary tissue assembly. Here, we systematically evaluate primary human intrahepatic cholangiocytes as a candidate population for such a platform and describe conditions that direct their branching morphogenesis. We find that extracellular matrix presentation, coupled with mitogen stimulation, promotes biliary branching in a Notch-dependent manner. These results demonstrate the utility of using 3D scaffolds for mechanistic investigation of cholangiocyte branching and provides a gateway to integrate biliary architecture in additionalin vitromodels of liver tissue.