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

gamma-Glutamyl transpeptidase (gamma GT) positive hepatocytes were isolated from F344 male rats fed 2-acetylaminofluorene. The isolation procedure is rapid and highly selective for cells exhibiting gamma GT 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-gamma GT antibody. gamma GT-positive cells bound to the dish within fifteen minutes and could be recovered as viable cells. Approximately 15% of the gamma GT-positive cells are isolated using this procedure. Novikoff hepatoma cells, a gamma GT-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-gamma GT antibody coated dishes was 50% inhibited by 2.8 mM sodium azide and 86% inhibited by 4.6 mM.