A rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects

Computational and Systems Biology Program, Massachusetts Institute of Technology, Cambridge, MA, USA.
DNA repair (Impact Factor: 3.36). 11/2011; 11(1):92-8. DOI: 10.1016/j.dnarep.2011.11.002
Source: PubMed

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.


Available from: Leona D Samson, Jun 02, 2015
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