A rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects
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Samson_A rapid survival.pdf
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Author(s)
Valiathan, Chandni
McFaline, Jose Luis
Samson, Leona D.
Date Issued
November 2011
Journal
DNA Repair
Publisher
Elsevier
Citation
Valiathan, Chandni, Jose L. McFaline, and Leona D. Samson. “A Rapid Survival Assay to Measure Drug-Induced Cytotoxicity and Cell Cycle Effects.” DNA Repair 11, no. 1 (January 2012): 92–98.
Version
Author's final manuscript
Abstract
We describe a rapid method to accurately measure the cytotoxicity of mammalian cells upon exposure to various drugs. Using this assay, we obtain survival data in a fraction of the time required to perform the traditional clonogenic survival assay, considered the gold standard. The dynamic range of the assay allows sensitivity measurements on a multi-log scale allowing better resolution of comparative sensitivities. Moreover, the results obtained contain additional information on cell cycle effects of the drug treatment. Cell survival is obtained from a quantitative comparison of proliferation between drug-treated and untreated cells. During the assay, cells are treated with a drug and, following a recovery period, allowed to proliferate in the presence of bromodeoxyuridine (BrdU). Cells that synthesize DNA in the presence of BrdU exhibit quenched Hoechst fluorescence, easily detected by flow cytometry; quenching is used to determine relative proliferation in treated vs. untreated cells. Finally, this assay can be used in high-throughput format to simultaneously screen multiple cell lines and drugs for accurate measurements of cell survival and cell cycle effects after drug treatment.
MIT Department
Massachusetts Institute of Technology. Center for Environmental Health Sciences
Massachusetts Institute of Technology. Computational and Systems Biology Program
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Biology
Koch Institute for Integrative Cancer Research at MIT
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DOI of Published Version
https://doi.org/10.1016/j.dnarep.2011.11.002
