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An in vitro micronucleus assay for determining the radiosensitivity of hepatocytes.

An in vitro micronucleus assay was developed for primary cultures of rat hepatocytes and utilized to determine the oxygen enhancement ratio (OER). Freshly isolated Fischer 344 female rat hepatocytes were irradiated in suspension either in air or in anoxia, cultured for 60 h to allow for the maximum expression of micronuclei, fixed with methanol-glacial acetic acid, stained with the fluorescent dye 4',6-diamidino-2-phenylindole (DAPI), and scored with a fluorescent microscope for the presence of micronuclei. The percentage of cells with micronuclei increased linearly with dose, and the slopes of the relationships were 0.044 +/- 0.001 Gy-1 and 0.015 +/- 0.001 Gy-1 in air and anoxia, respectively. The calculated OER of 2.9 +/- 0.5 is similar to that previously obtained for hepatocyte cell survival. Our data demonstrate that this in vitro hepatocyte micronucleus assay is a rapid and sensitive method to further investigate those factors which influence the radiosensitivity of hepatocytes.