Protocol for generation of human gallbladder epithelia cells and their derived hepatocytes using a chemically defined approach

The large-scale production of functional hepatocytes is critical for liver disease treatment, disease modeling, and drug development. Here, we present a protocol for isolating, expanding, and maintaining human gallbladder epithelial cells (hGBECs). We describe steps for characterizing hGBECs and their differentiation into hepatocytes. We then detail procedures to assess the functionality of the derived hepatocytes using key hepatic function assays. For complete details on the use and execution of this protocol, please refer to Chen et al.1.