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Identifying DNA Mutations in Purified Hematopoietic Stem/Progenitor Cells
Published on: February 24, 2014
In vitro mouse lymphoma (L5178Y Tk⁺/⁻-3.7.2C) forward mutation assay
Melissa R Schisler1, Martha M Moore, B Bhaskar Gollapudi
1The Dow Chemical Company, Toxicology & Environmental Research & Consulting, Midland, MI, USA.
Methods in Molecular Biology (Clifton, N.J.)
|July 31, 2013
Summary
The mouse lymphoma assay (MLA) is a key genetic toxicology test detecting mutations. It identifies various genetic events, aiding in chemical safety assessments.
Area of Science:
- Genetic Toxicology
- In Vitro Assays
Background:
- The mouse lymphoma assay (MLA) is a widely used method in genetic toxicology.
- It detects forward mutations at the thymidine kinase (Tk) locus in the L5178Y cell line.
Purpose of the Study:
- To detail the methodology and applications of the mouse lymphoma assay.
- To highlight its capability in detecting diverse genetic alterations.
Main Methods:
- Utilizes the L5178Y (Tk (+/-) -3.7.2C) mouse lymphoma cell line.
- Employs two cloning methods: soft agar or 96-microwell liquid media.
- Distinguishes between small- and large-colony mutants.
Main Results:
- Detects point mutations, deletions, chromosomal rearrangements, mitotic recombination, and nondisjunction.
- Small-colony mutants correlate with chromosomal aberrations; large-colony mutants with point mutations.
- Assay performance is influenced by cell line source, karyotype, and culture conditions.
Conclusions:
- The MLA provides valuable genotoxicity hazard information.
- Standardized MLA performance is crucial for accurate safety assessment of test substances.
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01:16In-vitro Mutagenesis
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.