Comparison of different cytotoxicity measurements for the in vitro micronucleus assay using L5178Y and TK6 cells in support of OECD draft Test Guideline 487.

The reference genotoxic agents mitomycin C, cadmium chloride, 2-aminoanthracene, vinblastine sulphate and 5-fluorouracil were tested in the in vitro micronucleus assay, in mouse lymphoma L5178Y cells and in human lymphoblastoid cells TK6, without cytokinesis block. This was done in support of the toxicity measures recommended in the late 2007 version of the draft OECD Test Guideline 487 for the testing of chemicals. Relative Population Doubling and Relative Increase in Cell Counts, used for the selection of the highest concentrations to be evaluated for genotoxicity assessment, based on a 50±5% cytotoxicity, both allowed to equally detect positive mitomycin C, cadmium chloride, 2-aminoanthracene, vinblastine sulphate and 5-fluorouracil on L5178Y and/or TK6 cells. Therefore, these parameters, recommended in the draft Test Guideline 487, are suitable to select the concentrations at the cytotoxicity required for genotoxicity assessment in the in vitro micronucleus assay without cytokinesis block.

[1]  Sac-fry Stages,et al.  OECD GUIDELINE FOR TESTING OF CHEMICALS , 2002 .

[2]  M. Fenech,et al.  HUMN project: detailed description of the scoring criteria for the cytokinesis-block micronucleus assay using isolated human lymphocyte cultures. , 2003, Mutation research.

[3]  A. Elhajouji,et al.  Mitomycin C, 5-fluoruracil, colchicine and etoposide tested in the in vitro mammalian cell micronucleus test (MNvit) in the human lymphoblastoid cell line TK6 at Novartis in support of OECD draft Test Guideline 487. , 2010, Mutation research.

[4]  H. Stopper,et al.  Evaluation of the in vitro micronucleus test as an alternative to the in vitro chromosomal aberration assay: position of the GUM Working Group on the in vitro micronucleus test. Gesellschaft für Umwelt-Mutations-forschung. , 1998, Mutation research.

[5]  David Kirkland,et al.  Comparison of different methods for an accurate assessment of cytotoxicity in the in vitro micronucleus test. II: Practical aspects with toxic agents. , 2008, Mutation research.

[6]  M. Fellows,et al.  Cytotoxicity in cultured mammalian cells is a function of the method used to estimate it. , 2007, Mutagenesis.

[7]  S Albertini,et al.  Comparison of different methods for an accurate assessment of cytotoxicity in the in vitro micronucleus test. I. Theoretical aspects. , 2008, Mutation research.

[8]  Sheila M Galloway,et al.  Population doubling: A simple and more accurate estimation of cell growth suppression in the in vitro assay for chromosomal aberrations that reduces irrelevant positive results , 2004, Environmental and molecular mutagenesis.

[9]  David Kirkland,et al.  Evaluation of different cytotoxic and cytostatic measures for the in vitro micronucleus test (MNVit): introduction to the collaborative trial. , 2010, Mutation research.

[10]  W. Muster,et al.  Mouse lymphoma thymidine kinase locus gene mutation assay: International Workshop on Genotoxicity Test Procedures Workgroup report , 2000, Environmental and molecular mutagenesis.