Development and validation of a high-content screening in vitro micronucleus assay in CHO-k1 and HepG2 cells

Westerink, Walter M.A.; Schirris, Tom J.J.; Horbach, G. Jean; Schoonen, W.G.E.J.

doi:10.1016/j.mrgentox.2011.05.007

Cited by 64 publications

(48 citation statements)

References 27 publications

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“…Hep G2 cells have been widely employed to determine chromosomal damage induced by genotoxic agents and complex mixtures [32,33]. In addition, it has been reported to be successfully used to discriminate aneugens and clastogens with signature of micronucleus size in a high-throughput screening system [34]. In combination with the flow cytometry-based MN test with Hep G2, the centralized source waters in this study were designated as either non-genotoxic or aneugenic or clastogenic, which indicates that along with the MN counts the information on genotoxic modes of action was simultaneously acquired in the present analytical platform.…”

Section: Discussionmentioning

confidence: 99%

Assessing of genotoxicity of 16 centralized source-waters in China by means of the SOS/umu assay and the micronucleus test: Initial identification of the potential genotoxicants by use of a GC/MS method and the QSAR Toolbox 3.0

Yan

Jiang²,

Na³

et al. 2014

Mutation Research/Genetic Toxicology and Environmental Mutagene

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Only few studies were conducted to assess genotoxicity of centralized source waters in China and almost none of them dealt with the causal relationship between the genotoxic effect and genotoxicants. In this work, 16 centralized source waters in China were sampled from five river systems and genotoxicity of their organic extracts was assessed by use of the SOS/umu test for DNA-damaging effect and the miniaturized flow cytometry-based micronucleus (MN) test for chromosome-damaging effect. In addition, initial identification of potential genotoxicants for the six samples from the Yangtze River was done with a GC/MS method and the QSAR toolbox 3.0. The results demonstrate that eight samples showed both indirect and direct DNA-damaging effects, another four samples showed only indirect DNA-damaging effects, while chromosome-damaging effects were found for 14 out of the 16 samples, in which aneugenic and clastogenic modes of action were found for 4 and 10 samples, respectively. Both direct/indirect DNAdamaging effects and chromosome-damaging effects were induced by the six Yangtze River samples, and the existing different types of genotoxicant confirmed the results. Furthermore, o-phenylphenol was initially identified as the major cause for the DNA-damaging effects while PAHs, pesticides, phenol and anthraquinone were identified as ubiquitous chromosome-damaging agents among these samples. In conclusion, a combination of the SOS/umu test and the miniaturized flow cytometry-based MN test to detect both DNA-damaging and chromosome-damaging effects could be used as a comprehensive genotoxicity assessment tool for the evaluation and classification of genotoxicity of complex mixtures, and potential genotoxicants can be initially identified with additional information from chemical analysis and the QSAR toolbox.

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Section: Discussionmentioning

confidence: 99%

Assessing of genotoxicity of 16 centralized source-waters in China by means of the SOS/umu assay and the micronucleus test: Initial identification of the potential genotoxicants by use of a GC/MS method and the QSAR Toolbox 3.0

Yan

Jiang²,

Na³

et al. 2014

Mutation Research/Genetic Toxicology and Environmental Mutagene

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“…Cells treated with test compound are then grown in the presence of cytochalasin-B to prevent the cytoplasmic division after nuclear division, fixed, stained and scored for binucleated and micronucleated cells (Fenech, 2000;SanchezLamar et al, 2002). The detection of micronuclei, manual or, as more recently described, automated (Diaz et al, 2007;Westerink et al, 2011), provides a readily measurable index of chromosome breakage and loss. Although the test is unable to provide a measure of more subtle changes, such as balanced translocations, such a level of detail is in most cases unnecessary (Fenech, 1997).…”

Section: In Vitro Methods For Genotoxicity Assessmentmentioning

confidence: 99%

Review of current and “omics” methods for assessing the toxicity (genotoxicity, teratogenicity and nephrotoxicity) of herbal medicines and mushrooms

Ouédraogo

Baudoux

Stévigny

et al. 2012

Journal of Ethnopharmacology

124

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“…The CBMN assay was conducted according to Yan et al (2014a) and Westerink et al (2011). Briefly, a complete culture medium was prepared with 90% F12 medium, 10% fetal calf serum, and 1% penicillin-streptomycin (10,000 U/mL, HyClone).…”

Section: Cbmn Assaymentioning

confidence: 99%

“…Recently, a highcontent screening (HCS) technique was developed in the rodent cell line CHO-K1 that complements the CBMN assay by automatically analyzing cells treated with fluorescent dyes in microplates (Diaz et al, 2007;Ogata et al, 2011;Westerink et al, 2011). The HCS provides several key advantages over the traditional method used with the bioassay.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of genotoxic effects of surface waters using a battery of bioassays indicating different mode of action

Han

Oda

et al. 2016

Ecotoxicology and Environmental Safety

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a b s t r a c tWith the burgeoning contamination of surface waters threatening human health, the genotoxic effects of surface waters have received much attention. Because mutagenic and carcinogenic compounds in water cause tumors by different mechanisms, a battery of bioassays that each indicate a different mode of action (MOA) is required to evaluate the genotoxic effects of contaminants in water samples. In this study, 15 water samples from two source water reservoirs and surrounding rivers in Shijiazhuang city of China were evaluated for genotoxic effects. Target chemical analyses of 14 genotoxic pollutants were performed according to the Environmental quality standards for surface water of China. Then, the in vitro cytokinesis-block micronucleus (CBMN) assay, based on a high-content screening technique, was used to detect the effect of chromosome damage. The SOS/umu test using strain TA1535/pSK1002 was used to detect effects on SOS repair of gene expression. Additionally, two other strains, NM2009 and NM3009, which are highly sensitive to aromatic amines and nitroarenes, respectively, were used in the SOS/umu test to avoid false negative results. In the water samples, only two of the genotoxic chemicals listed in the water standards were detected in a few samples, with concentrations that were below water quality standards. However, positive results for the CBMN assay were observed in two river samples, and positive results for the induction of umuC gene expression in TA1535/pSK1002 were observed in seven river samples. Moreover, positive results were observed for NM2009 with S9 and NM3009 without S9 in some samples that had negative results using the strain TA1535/pSK1002. Based on the results with NM2009 and NM3009, some unknown or undetected aromatic amines and nitroarenes were likely in the source water reservoirs and the surrounding rivers. Furthermore, these compounds were most likely the causative pollutants for the genotoxic effect of these water samples. Therefore, to identify causative pollutants with harmful biological effects, chemical analyses for the pollutants listed in water quality standards is not sufficient, and single-endpoint bioassays may underestimate adverse effects. Thus, a battery of bioassays based on different MOAs is required for the comprehensive detection of harmful biological effects. In conclusion, for genotoxicity screening of surface waters, the SOS/umu test system by using different strains combined with the CBMN assay was a useful approach.

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Development and validation of a high-content screening in vitro micronucleus assay in CHO-k1 and HepG2 cells

Cited by 64 publications

References 27 publications

Assessing of genotoxicity of 16 centralized source-waters in China by means of the SOS/umu assay and the micronucleus test: Initial identification of the potential genotoxicants by use of a GC/MS method and the QSAR Toolbox 3.0

Assessing of genotoxicity of 16 centralized source-waters in China by means of the SOS/umu assay and the micronucleus test: Initial identification of the potential genotoxicants by use of a GC/MS method and the QSAR Toolbox 3.0

Review of current and “omics” methods for assessing the toxicity (genotoxicity, teratogenicity and nephrotoxicity) of herbal medicines and mushrooms

Evaluation of genotoxic effects of surface waters using a battery of bioassays indicating different mode of action

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