Publisher Summary This chapter discusses preparation of isolated rat liver cells The early mechanical and chemical methods for liver-cell preparation were relatively successful in converting liver tissue to a suspension of isolated cells The successful preparation of intact liver cells by perfusion with collagenase is technically quite difficult The major method for preparation of nonparenchymal liver cells is based on the selective sensitivity of parenchymal cells toward proteases By incubation of rat liver minces with pronase, most of the liver parenchyma is digested, while nonparenchymal cells remain intact and can be recovered from the incubate Similar results have been reported with trypsin digestion of collagenase-dispersed liver minces, but pronase appears to be more effective The most common procedure is to perfuse the liver briefly with pronase before it is minced and incubated with the enzyme Such direct pronase methods have been used by several investigators with yields of nonparenchymal liver cells reported to be in the range 2–15 × 10 6 cells/gm liver

Preparation of isolated rat liver cells.

Publisher Summary Glutathione (GSH) is the most important nonprotein thiol in living systems and is of widespread occurrence in the intracellular milieu of animals, plants, and microorganisms GSH was isolated and named by the English biochemist Frederick Gowland Hopkins This chapter discusses GSH status, the biologically relevant chemistry of GSH, the forms in which GSH can be present within the cell, along with the GSH content of cells and the methods for analysis of this substance GSH-related biochemical reactions and the biological roles of GSH are discussed in the chapter The use of perturbations in GSH status as a means for investigating GSH-related phenomena and an analysis of the consequences of perturbation are presented A short summary of genetic lesions related to GSH is also included Like chemically induced perturbations in GSH status, genetic lesions provide valuable insights into the role of GSH in normal functions and processes in cells The chapter concludes with some brief comments about the future of the relationship of GSH status to cellular processes

The Glutathione Status of Cells

12 – Lipid Peroxidation: Mechanisms, Analysis, Enzymology and Biological Relevance

Glutathione metabolism and its role in hepatotoxicity.

In vivo and in vitro uptake and degradation of acetylated low density lipoprotein by rat liver endothelial, Kupffer, and parenchymal cells.

Distribution of L- and M-type pyruvate kinase between parenchymal and Kupffer cells of rat liver

Two interconvertible forms of L-type pyruvate kinase from rat liver

On the lipid peroxidation of rat liver hepatocytes, the formation of fluorescent chromolipids and high molecular weight protein.

On the molecular basis of pyruvate kinase deficiency. I. Primary defect or conseqence of increased glutathione disulfide concentration.

M-type pyruvate kinase of leucocytes: An allosteric enzyme

Th.J.C. Van Berkel

Papers

Distribution of L- and M-type pyruvate kinase between parenchymal and Kupffer cells of rat liver

Two interconvertible forms of L-type pyruvate kinase from rat liver

On the lipid peroxidation of rat liver hepatocytes, the formation of fluorescent chromolipids and high molecular weight protein.

On the molecular basis of pyruvate kinase deficiency. I. Primary defect or conseqence of increased glutathione disulfide concentration.

M-type pyruvate kinase of leucocytes: An allosteric enzyme