An assessment of the in vivo rat hepatocyte DNA-repair assay

Activity of 2-Acetylaminofluorene as a UDS inducting agent in B6C3F1 mouse hepatocytes in vitro

“…This effect has been recorded before in this and other laboratories for a variety of compounds using isolated hepatocytes from several species (eg. Bermudez et al, 1979;McQueen and Williams, 1983;Kornbrust and Barfknecht, 1984;Ashby et al, 1985;Steinmetz et al, 1988) and emphasizes the need for careful selection of dose levels in such assays.…”

“…Animals were dosed by oral gavage with corn oil (Kraft-Wesson) at 10 ml/kg 12 h before sacrifice as part of an in vivo/in vitro study. Hepatocytes were isolated by a two-stage liver perfusion system (Seglen, 1976;Ashby et al, 1985) using the Krebs-Ringer phosphate and Krebs-Ringer hydrogen carbonate buffers described by Mitchell et al (1984). Rat hepatocytes were isolated as described by Ashby et al (1985): mouse hepatocytes were prepared by the methods previously described (Lefevre and Ashby, 1989).…”

“…Cells were fixed, washed, mounted, autoradiographed and processed as described by Ashby et al (1985). Hepatocytes were assessed for UDS using an automated counting system .…”

“…This chemical is a potent inducer of UDS in isolated rat hepatocytes (Williams 1977;Ashby et al, 1985) and in the in vivo/in vitro rat liver UDS assay (Mirsalis and Butterworth, 1980. However, it gives a generally negative profile in mouse hepatocytes.…”

Genotoxicity testing of esterified propoxylated glycerol (EPG)

“…Three independent experiments, a range-finding and two main studies (Experiments 1 and 2), were conducted according to the procedure detailed by Ashby et al (1985) as modified from the original assay developed by Mirsalis and Butterworth (1980) to assess the potential of EPG (H-EPG-05 HR/SO 9:1; H-EPG-05 soyate; H-EPG-14 soyate) to induce unscheduled DNA synthesis (UDS) in liver cells isolated from rats exposed orally.…”