4] Isolation and use of liver cells

Abstract

This chapter describes the isolation and use of liver cells. The technique is based on liver perfusion with collagenase after removal of Ca 2+ by preperfusion with a chelator. Moderately high yields of viable, single hepatocytes with a smooth, spherical appearance and essentially free of nonparenchymal cells are obtained. Early attempts to isolate hepatocytes employed mechanical force and, subsequently, perfusion of the liver with Ca 2+- or K+-chelators but were unsuccessful in obtaining viable cells in high yields. This was not achieved until the isolation of rat hepatocytes by the use of the digestive enzymes collagenase and hyaluronidase was introduced by Howard and collaborators. Although most of the techniques recently employed to obtain isolated hepatocytes involve perfusion of the liver with digestive enzymes, attempts are also made to avoid the perfusion step and achieve hepatocyte isolation by incubating cut pieces of the liver in enzyme-containing solutions. The presently available results suggest that isolated hepatocytes—and cell suspensions isolated from certain other target tissues—may be a useful model for drug toxicity studies in the future.