Protocol for modeling biliary cell activation in human precision-cut liver slices

: Persistent ductular reaction is a hallmark of chronic liver injury, characterized by the proliferation and pro-inflammatory activity of reactive biliary epithelial cells. Here, we present a protocol for modeling biliary cell activation in human precision-cut liver slices. We describe steps for collecting samples, generating, and culturing precision-cut liver slices, and activating biliary cells ex vivo while preserving native architecture and multicellular interactions. Biliary cell activation can be quantified by EdU incorporation, supporting mechanistic studies and translational drug-screening applications. For complete details on the use and execution of this protocol, please refer to Anfuso et al.1.