Isolation and Culture of Primary Hepatocytes from Resected Human Liver Tissue

As our knowledge of the species differences in drug metabolism and drug-induced hepatotoxicity has expanded significantly, the need for human-relevant in vitro hepatic model systems has become more apparent than ever before. Human hepatocytes have become the "gold standard" for evaluating hepatic metabolism and toxicity of drugs and other xenobiotics in vitro. In addition, they are becoming utilized more extensively for many kinds of biomedical research, including a variety of biological, pharmacological, and toxicological studies. This chapter describes methods for the isolation of primary human hepatocytes from liver tissue obtained from an encapsulated end wedge removed from patients undergoing resection for removal of liver tumors or from resected segments from whole livers obtained from multi-organ donors. In addition, methods are described for culturing primary hepatocytes under various matrix compositions and geometries, which reestablish intercellular contacts and normal cellular architecture for optimal phenotypic gene expression and response to drugs and other xenobiotics in vitro. Overall, improved isolation, cultivation, and preservation methods have expanded the number of applications for primary human hepatocytes in basic research, which has allowed for exciting advances in our understanding of the biochemical and molecular mechanisms of human liver toxicity and disease.