Isolation of Hepatic Stellate Cells and Lymphocytes for Co-culture Systems

In contrast to quiescent hepatic stellate cells (HSCs), activated HSCs play crucial roles in the development of liver fibrosis by producing a huge amount of extracellular matrix such as collagen fibers. However, recent lines of evidence have also highlighted the immunoregulatory functions of HSCs, in which they interact with diverse hepatic lymphocytes to produce cytokines and chemokines, release extracellular vesicles, or express specific ligands. Therefore, to understand the exact interactions between HSCs and lymphocyte subsets in the pathogenesis of the liver disease, it is valuable to establish experimental procedures to isolate HSC and co-culture them with lymphocytes. Here, we introduce the efficient methods to isolate and purify mouse HSCs and hepatic lymphocytes using density gradient centrifugation, microscopic observation, and flow cytometry. Moreover, we provide the direct and indirect co-culturing methods of isolated mouse HSCs and hepatic lymphocytes based upon the purpose of the study.