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In Vitro Mouse Lymphoma Cell (L5178Y Tk+/− −3.7.2.C) Forward Mutation Assay

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Genotoxicity Assessment

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2031))

Abstract

The in vitro mouse lymphoma cell assay (MLA) is one of the most widely practiced assays in genetic toxicology. MLA detects forward mutations at the thymidine kinase (Tk) locus of the L5178Y (Tk+/− −3.7.2C) cell line derived from a mouse thymic lymphoma. This assay is capable of detecting a wide range of genetic events including point mutations, deletions and multilocus, chromosomal rearrangements, mitotic recombination and nondisjunction. There are two equally accepted versions of the assay, one using soft agar cloning and the second method using liquid media cloning in 96-microwell plates. There are two morphologically distinct types of mutant colonies recovered in the MLA; small and large colony mutants. The induction of small colony mutants is associated with chemicals inducing gross chromosomal aberrations, whereas the induction of large mutant colonies is generally associated with chemicals inducing point mutations. The source and karyotype of the cell line as well as the culture conditions are important variables that could influence the assay performance. The assay when performed according to the standards recommended by the International Workshops on Genotoxicity Testing (IWGT) and the Organization of Economic Cooperation and Development Test Guideline 490 is capable of providing valuable genotoxicity hazard information as part of the overall safety assessment process of various classes of test substances.

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Author information

Authors and Affiliations

  1. Environmental, Health & Safety, The Dow Chemical Company, Midland, MI, USA

    Melissa R. Schisler

  2. Ramboll, Little Rock, AR, USA

    Martha M. Moore

  3. Exponent Engineering and Scientific Consulting, Alexandria, VA, USA

    B. Bhaskar Gollapudi

Authors
  1. Melissa R. Schisler
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  2. Martha M. Moore
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  3. B. Bhaskar Gollapudi
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Corresponding author

Correspondence to Melissa R. Schisler .

Editor information

Editors and Affiliations

  1. CSIR—Indian Institute of Toxicology Research, Lucknow, Uttar Pradesh, India

    Alok Dhawan

  2. New Delhi, Delhi, India

    Mahima Bajpayee

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Schisler, M.R., Moore, M.M., Gollapudi, B.B. (2019). In Vitro Mouse Lymphoma Cell (L5178Y Tk+/− −3.7.2.C) Forward Mutation Assay. In: Dhawan, A., Bajpayee, M. (eds) Genotoxicity Assessment. Methods in Molecular Biology, vol 2031. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9646-9_1

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  • DOI: https://doi.org/10.1007/978-1-4939-9646-9_1

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9645-2

  • Online ISBN: 978-1-4939-9646-9

  • eBook Packages: Springer Protocols

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