Programa del congreso

Engineering functional livers is critical to alleviate the global shortage of donor organs. Reconstituting an organized biliary tree within a bioengineered liver remains a major challenge, requiring a cell source capable of large-scale expansion and precise differentiation. Here, we demonstrate that human intrahepatic cholangiocyte organoids (ICOs) can reconstruct a functional biliary network within decellularized rat liver scaffolds. ICOs derived from fresh or cryopreserved hepatocytes were expanded in spinner flasks, dissociated, and delivered via the bile duct using an alternating high- and low-pressure injection strategy to ensure coverage of large and small intrahepatic ducts. Following 7 days of perfusion culture, ICOs adhered to the scaffold, displayed reduced Ki67 expression, and maintained ductal compartmentalization without parenchymal invasion. High-pressure pulses promoted continuous biliary-like structures spanning multiple ductal levels. Immunofluorescence confirmed cholangiocyte-specific markers, and mature gene expression (CK19, CK7) was upregulated. This approach establishes ICOs as a scalable cell source for biliary reconstruction and introduces a pressure-controlled seeding method that preserves anatomical fidelity. Our findings represent a critical step toward generating bioengineered livers with functional biliary networks and support translation to large-animal models for transplantation applications.