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Internal standards for survival: increasing the accuracy for human cell mutation assays
Mutation Research
|April 1, 1987
Summary
A novel internal standard method corrects bias in mutation assays caused by varying cell densities. This approach improves the accuracy of determining mutant fractions, especially for rare variants in cell transformation studies.
Area of Science:
- Toxicology
- Genetics
- Cell Biology
Background:
- Conventional mutation assays often use varying cell densities, introducing bias in mutant fraction determination.
- Higher initial cell densities can affect colony formation, impacting assay accuracy.
Purpose of the Study:
- To introduce a reliable method for accurate mutant fraction determination in cell mutation assays.
- To address the bias caused by cell density variations in mutation and cell transformation assays.
Main Methods:
- Inclusion of an experimentally distinguishable internal standard cell population.
- Utilizing high cell density plating for enhanced detection of rare variants.
- Measuring mutation to 6-thioguanine resistance in a human B lymphoblast line.
Main Results:
- The internal standard method demonstrated high satisfaction in mutation assays.
- Calculated induced mutant fractions were significantly lower using the internal-standard method compared to conventional methods.
- The method proved effective for treatments with anti-2,3-dihydroxy-1,10b-epoxy-1,2,3-trihydrofluoranthene (FDE), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), and 4-nitroquinoline-oxide (4NQO).
Conclusions:
- The internal standard method provides a necessary feature for accurate rare variant fraction determination.
- This approach is applicable to all single cell mutation or transformation assays.
- The proposed method enhances the reliability of mutation detection in toxicological and genetic studies.