Use of rat primary lung cells for studying genotoxicity with the sister-chromatid exchange and micronucleus assays.
Whong-Z; Stewart-JD; Ong-T
Mutat Res 1990 May; 241(1):7-13
To evaluate pulmonary genotoxicity both sister chromatid exchange (SCE) and micronucleus (MN) genotoxicity assays were characterized in the primary lung cells of 6 to 8 week old male CD-rats. Several enzyme systems used to isolate lung cells were compared. The method of cytokinesis block for identifying divided cells was established in the in-vitro MN test. The highest yield of viable and colony forming cells in the cell isolation evaluation was obtained with a combined enzyme separation of rat lungs with trypsin (1.3 milligrams/milliliter), plus collagenase (50 units/milliliter). Treatment of primary lung cells with 2 micrograms cytochalasin (CYB)/milliliter for two days appeared optimal for MN scoring in CYB induced binucleated (BN) cells. A dose related increase in micronucleated BN cells in-vitro was obtained with this procedure by exposure to mitomycin-C (50077) (MMC), triethylenemelamine (51183), and benzo(a)pyrene (50328) without metabolic activation. Maximum second division metaphases were obtained in the SCE assay after the cells were incubated with bromodeoxyuridine (59143) for 48 to 54 hours. Also observed after this incubation period were high frequencies of SCE induced by MMC and 3-methylcholanthrene (56495), in-vitro and in-vivo with no S9 activation. The authors conclude that rat primary lung cells can metabolize polycyclic aromatic hydrocarbons and that this lung cell system is potentially useful for detecting pulmonary genotoxicants.
NIOSH-Author; Chromosome-damage; Chromosome-disorders; Genotoxic-effects; Cell-damage; Cell-function; Metabolic-study; Lung-cells;
Author Keywords: Rat primary lung cells; Sister-chromatid exchange; Micronucleus; Pulmonary genotoxicity studies
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