Centrifugal elutriation of hepatocytes from 2-acetylaminofluorene-treated rats and their characterization by flow cytometry

Treatment of male Wistar rats with 2-acetylaminofluorene (2-AAF) markedly altered the ploidy distribution of liver cells. Small diploid hepatocytes first appeared after 4-5 weeks feeding of a diet containing 0.02% 2-AAF; after 9 weeks 65-70% of the hepatocytes were diploid. Approximately two-thirds of this new liver cell population persisted after termination of the treatment. The hepatocytes from 2-AAF treated animals were separated according to size and ploidy by centrifugal elutriation and stained for γ-glutamyltrans-peptidase (γ-GTase). The percentage of γ-GTase-positive hepatocytes did not significantly differ between the various elutriated cell fractions. Thus γ-GTase-positive liver cells obtained by feeding of 2-AAF do not represent a distinct size class of hepatocytes. The significance of carcinogen-induced diploid hepatocytes in hepatocarcinogenesis is discussed.