- Analysis of the interplay between hepatitis B virus-positive hepatocytes and Kupffer cells ex vivo using mice as a model.
Analysis of the interplay between hepatitis B virus-positive hepatocytes and Kupffer cells ex vivo using mice as a model.
STAR protocols (2022-05-17)
Yumei Li, Jiyoung Lee, Doumet Georges Helou, Omid Akbari, Jing-Hsiung James Ou
PMID35573478
ABSTRACT
Kupffer cells play critical roles in both hepatitis B virus (HBV) persistence and clearance. Here, we provide a protocol for studying the interplay between Kupffer cells and HBV-positive hepatocytes ex vivo using mice as a model. This protocol includes hydrodynamic injection of HBV DNA into mouse hepatocytes, liver perfusion for isolating hepatocytes and Kupffer cells, and Seahorse metabolic analysis of Kupffer cells. This protocol allows the detailed analysis of how HBV-positive hepatocytes and Kupffer cells impact each other ex vivo. For complete details on the use and execution of this protocol, please refer to Li et al. (2022).
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
hEGF, EGF, recombinant, expressed in E. coli, lyophilized powder, suitable for cell culture
Sigma-Aldrich
Collagenase from Clostridium histolyticum, suitable for release of physiologically active rat hepatocytes, Type IV, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid
Sigma-Aldrich
Deoxyribonuclease I from bovine pancreas, lyophilized powder, Protein ≥85 %, ≥400 Kunitz units/mg protein