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Mutagenesis, Vol. 14, No. 4, 403-410, July 1999
© 1999 UK Environmental Mutagen Society/Oxford University Press

A micromethod for the in vitro micronucleus assay

Fabrice Nesslany1 and Daniel Marzin1,2,3

1 Laboratoire de Toxicologie Génétique, Institut Pasteur de Lille, 59019 Lille Cedex and 2 Département Toxicologie–Hydrologie–Hygiène, Université de Lille II, Droit et Santé, Faculté des Sciences Pharmaceutiques et Biologiques, 59006 Lille Cedex, France

A micromethod for the in vitro micronucleus assay was developed using L5178Y cells to enable the rapid screening of a large number of molecules. The method is quick, simple to perform and needs very small amounts of compound, i.e. <10 mg. In this methodology, three types of treatment were carried out in parallel, enabling an optimal detection of both aneugenic and clastogenic compounds: two treatments without metabolic activation with or without a recovery period after a 24 h continuous treatment and one treatment with metabolic activation by Aroclor 1254-induced rat or hamster liver S9 mix. Seventeen known genotoxins (12 clastogens and five aneugens) and seven known non-genotoxins were tested. The in vitro micronucleus micromethod using L5178Y cells exhibited good sensitivity (16 positive/17 known genotoxins tested) and specificity (7 negative/7 known non-genotoxins tested) for the 24 test compounds studied with or without metabolic activation. Furthermore, this test showed a good correlation with other in vitro micronucleus tests performed using macromethods with various mammalian cell cultures. We conclude that the in vitro micronucleus micromethod with L5178Y cells could be used in the earliest stages of development of new molecules as a preliminary short-term screening assay before starting regulatory tests.

3 To whom correspondence should be adressed at: Laboratoire de Toxicologie Génétique, Institut Pasteur de Lille, 1 rue du Professeur Calmette, 59019 Lille Cedex, France. Tel: +33 3 20 87 79 75; Fax: +33 3 20 87 73 10; Email: daniel.marzin{at}pasteur-lille.fr


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