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.
D000818:Animals; D001666:Binding Sites, Antibody; D002469:Cell Separation; D005260:Female; D006651:Histocytochemistry; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D008114:Liver Neoplasms, Experimental / Q000201:enzymology* / Q000473:pathology; D008297:Male; D011230:Precancerous Conditions / Q000201:enzymology* / Q000473:pathology; D051381:Rats; D011916:Rats, Inbred F344; D011920:Rats, Inbred WF; D005723:gamma-Glutamyltransferase / Q000032:analysis*
D005723:gamma-Glutamyltransferase