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Int. J. Mol. Sci. 2018, 19(1), 262; https://doi.org/10.3390/ijms19010262

Mutation Analysis in Cultured Cells of Transgenic Rodents

1
Department of Preventive Medicine, University of Southern California Keck School of Medicine, University of Southern California, M/C 9603, Los Angeles, CA 90033, USA
2
Department of Cancer Biology, Beckman Research Institute of the City of Hope, Duarte, CA 91010, USA
*
Author to whom correspondence should be addressed.
Received: 20 December 2017 / Revised: 10 January 2018 / Accepted: 15 January 2018 / Published: 16 January 2018
(This article belongs to the Section Biochemistry)
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Abstract

To comply with guiding principles for the ethical use of animals for experimental research, the field of mutation research has witnessed a shift of interest from large-scale in vivo animal experiments to small-sized in vitro studies. Mutation assays in cultured cells of transgenic rodents constitute, in many ways, viable alternatives to in vivo mutagenicity experiments in the corresponding animals. A variety of transgenic rodent cell culture models and mutation detection systems have been developed for mutagenicity testing of carcinogens. Of these, transgenic Big Blue® (Stratagene Corp., La Jolla, CA, USA, acquired by Agilent Technologies Inc., Santa Clara, CA, USA, BioReliance/Sigma-Aldrich Corp., Darmstadt, Germany) mouse embryonic fibroblasts and the λ Select cII Mutation Detection System have been used by many research groups to investigate the mutagenic effects of a wide range of chemical and/or physical carcinogens. Here, we review techniques and principles involved in preparation and culturing of Big Blue® mouse embryonic fibroblasts, treatment in vitro with chemical/physical agent(s) of interest, determination of the cII mutant frequency by the λ Select cII assay and establishment of the mutation spectrum by DNA sequencing. We describe various approaches for data analysis and interpretation of the results. Furthermore, we highlight representative studies in which the Big Blue® mouse cell culture model and the λ Select cII assay have been used for mutagenicity testing of diverse carcinogens. We delineate the advantages of this approach and discuss its limitations, while underscoring auxiliary methods, where applicable. View Full-Text
Keywords: assay; cII transgene; carcinogen; mouse embryonic fibroblasts; mutation assay; cII transgene; carcinogen; mouse embryonic fibroblasts; mutation
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Besaratinia, A.; Zheng, A.; Bates, S.E.; Tommasi, S. Mutation Analysis in Cultured Cells of Transgenic Rodents. Int. J. Mol. Sci. 2018, 19, 262.

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