Chapter 29 The Preparation and Characterization of Intact Isolated Parenchymal Cells from Rat Liver

The concept of utilizing isolated cell suspensions as a tool to study metabolic events in the liver in vitro has been attractive for many years. The advantages of such a system would be manyfold. Numerous attempts to obtain isolated parenchymal cells from adult rodent liver have been made over the past few decades. The early methods involved mechanical treatment of the tissue, alone or in combination with enzymic digestion, or perfusion of the intact liver with chelating agents, such as EDTA and citrate. It was not until 1967, however, that structurally and metabolically intact isolated parenchymal cells from adult rat liver could be obtained in sufficient yield to permit further characterization. This was achieved by perfusing the liver with, and incubating the sliced tissue in, a calcium-free medium containing collagenase and hyaluronidase under physiological conditions. This method, with more recent modifications is described in this chapter. In addition, the evaluation of the structural and metabolic integrity of these cells and problems encountered in using such a preparation are discussed.