Functional bile duct constructs can be created in vitro using EDO or BDO. (a–d) Acetylated α‐tubulin staining shows the presence of cilia in EDO and BDO recellularized samples. Scale bars: (a) 100 µm, (b–d): 50 µm. (e) TEER measurements show an increase in resistance compared to decellularized ECM (N = 3 measurements per sample). (f) and (g): Ussing chamber experiments showing short circuit current (I
sc) changes upon consecutive addition of Forskolin (a cAMP agonist), GlyH‐101 (CFTR inhibitor), and UTP (Ca2+ agonist and CaCC activator) of EDO recellularized ECM (f, N = 2) and BDO recellularized ECM (g, N = 2). After addition of Forskolin a small response in EDO samples was recorded, but not in the BDO samples. GlyH‐101 successfully blocked CFTR‐channel activity in both samples, as a decrease I
sc was recorded. Addition of 50 µM UTP showed an increase in current for both samples indicating the presence of CaCC. BDO, bile‐derived organoids; CaCC, calcium activated chloride channels; ECM, extracellular matrix; EDO, extrahepatic bile duct‐derived organoids; TEER, trans epithelial electrical resistance [Color figure can be viewed at wileyonlinelibrary.com]