Kinetochore localization in micronucleated cytokinesis-blocked Chinese hamster ovary cells: a new and rapid assay for identifying aneuploidy-inducing agents.

We have developed a modified micronucleus assay using an antikinetochore antibody and cytokinesis-blocked Chinese hamster ovary cells as a simple and rapid method for detecting aneuploidy-inducing agents. The presence of a kinetochore in a micronucleus of a binucleated cell indicates a cell with a high probability for aneuploidy following cytokinesis. The method requires minimal training to perform and score and can readily distinguish aneuploidy-inducing agents from clastogens. Micronucleated cells treated with the aneuploidy-inducing agents benomyl and vinblastine sulfate contained a kinetochore-positive micronucleus 92% and 94% of the time whereas micronucleated cells treated with the clastogen methyl methanesulfonate contained a kinetochore-positive micronucleus only 11% of the time. This relatively simple method for distinguishing aneuploidy-inducing agents from clastogenic agents may be used as a routine genotoxicity assay to identify environmental and therapeutic agents with aneuploidy-inducing properties.