Validation of a flow cytometric (FCM) in vitro rat hepatocyte DNA repair assay employing the bivariate BrdUrd-FITC/PI immunocytochemical technique

An in vitro FCM DNA repair assay has been developed. Cultures of rat hapatocytes were exposed to a battery of chemicals for 18-20 hrs. Compounds were selected based upon both their genotoxic and carcinogenic characteristics. Evidence of DNA repair was noted by detecting BrdUrd uptake. Low intensity...

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Bibliographic Details
Published inCytometry (Baltimore); (United States) Vol. 6
Main Authors Selden, J.R., Dolbeare, F., Clair, J.H., DeLuca, J.G.
Format Conference Proceeding
LanguageEnglish
Published United States 01.01.1993
Subjects
550300 > Cytology
550400 > Genetics
ANIMAL CELLS
AUTORADIOGRAPHY
BASIC BIOLOGICAL SCIENCES
BIOASSAY
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
COMPARATIVE EVALUATIONS
DNA REPAIR
EVALUATION
FLUORESCENCE
GENES
INDUCTION
LIVER CELLS
LUMINESCENCE
REPAIR
SOMATIC CELLS 550200 > Biochemistry
TOXICITY
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Summary:An in vitro FCM DNA repair assay has been developed. Cultures of rat hapatocytes were exposed to a battery of chemicals for 18-20 hrs. Compounds were selected based upon both their genotoxic and carcinogenic characteristics. Evidence of DNA repair was noted by detecting BrdUrd uptake. Low intensity BrdUrd-FITC fluorescent signals from repairing cells were visualized by use of linear uni- or bi-variate histograms. This assay's sensitivity was directly compared to that of autoradiography. Results revealed the following: (1) A high correlation exists between genotoxicity and DNA repair; (2) The results of these assays were generally in agreement; and, (3) The sensitivity of this FCM DNA repair assay compares favorably to that of autoradiography. Thus, this assay provides a sensitive and reliable means of identifying agents which induce DNA repair in mammalian cells.
Bibliography:None
CONF-9303114--
ISSN:0196-4763
1097-0320