Isolation of γ-glutamyl transpeptidase positive hepatocytes during the early stages of hepatocarcinogenesis in the rat

Abstract

γ-Glutamyl transpeptidase (γGT) positive hepatocytes were isolated from F344 male rats fed 2-acetylaminofluorene. The isolation procedure is rapid and highly selective for cells exhibiting γGT on their surface. Suspensions of liver cells obtained from perfusion in situ with a collagenase solution were incubated on Petri dishes coated with affinity purified rabbit anti-γGT antibody. γGT-positive cells bound to the dish within fifteen minutes and could be recovered as viable cells. Approximately 15% of the γGT-positive cells are isolated using this procedure. Novikoff hepatoma cells, a γGT-positive cell line, were used to define the parameters of the assay. The binding was both time and temperature dependent. Binding of cells to the anti-γGT antibody coated dishes was 50% inhibited by 2.8 mM sodium azide and 86% inhibited by 4.6 mM.