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Development and characterization of a stable epithelial cell line from Muta Mouse lung
Paul A White1, George R Douglas, John Gingerich
1Mutagenesis Section, Safe Environments Program, Health Canada, Ottawa, Ontario, Canada. paul_white@hc-sc.gc.ca
Environmental and Molecular Mutagenesis
|October 14, 2003
Summary
A new Muta Mouse lung epithelial cell line offers a stable in vitro model for mutagenicity testing. This cell line complements in vivo assays, providing a reliable system for evaluating genetic mutations with identical endpoints.
Area of Science:
- Toxicology and Mutagenesis
- In Vitro Toxicology
- Genetics and Genomics
Background:
- Existing in vitro mutagenicity assays often lack direct correlation with in vivo outcomes.
- Development of reliable cell lines is crucial for advancing predictive toxicology.
- The Muta Mouse system provides an in vivo model for genetic mutation detection.
Purpose of the Study:
- To isolate and characterize a stable epithelial cell line from Muta Mouse lung.
- To evaluate the cell line's suitability as an in vitro complement to the in vivo Muta Mouse assay.
- To assess the cell line's responsiveness to various mutagens and its utility in routine mutagenicity testing.
Main Methods:
- Isolation and characterization of a lung epithelial cell line.
- Genomic stability assessment across multiple generations.
- Measurement of spontaneous mutation rates (micronuclei, sister chromatid exchanges, chromosome aberrations).
- Analysis of transgene integration (lacZ loci, monomers) using FISH and slot-blot.
- Mutagenicity testing with direct-acting mutagens (N-ethyl-N-nitrosourea, ICR-191), a polycyclic aromatic hydrocarbon (Benzo[a]pyrene), and a heterocyclic amine (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine) with and without metabolic activation (rat liver S9).
- Assessment of xenobiotic resistance related to glutathione-S-transferase activity.
Main Results:
- A stable, contact-inhibited epithelial cell line with retained pulmonary characteristics was established.
- The cell line demonstrated stable genome and low spontaneous rates of genetic damage.
- Consistent spontaneous mutant frequency at the lacZ transgene was observed.
- The cell line responded to mutagens, with Benzo[a]pyrene inducing significant mutation frequency without exogenous metabolic activation, while 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine required rat liver S9.
- High glutathione-S-transferase activity correlated with resistance to cytotoxic effects.
Conclusions:
- The developed Muta Mouse lung epithelial cell line serves as a valuable in vitro model for mutagenicity testing.
- It effectively complements the in vivo Muta Mouse assay by utilizing identical genetic endpoints.
- This cell line facilitates routine in vitro mutagenicity assessments, enhancing toxicological evaluations.