Title | A rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects. |
Publication Type | Journal Article |
Year of Publication | 2012 |
Authors | Valiathan, C, McFaline, JL, Samson, LD |
Journal | DNA Repair (Amst) |
Volume | 11 |
Issue | 1 |
Pagination | 92-8 |
Date Published | 2012 Jan 2 |
ISSN | 1568-7856 |
Keywords | Biological Assay, Carmustine, Cell Adhesion, Cell Cycle, Cell Death, Cell Line, Cell Proliferation, Cell Survival, Humans |
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. |
DOI | 10.1016/j.dnarep.2011.11.002 |
Alternate Journal | DNA Repair (Amst.) |
PubMed ID | 22133811 |
PubMed Central ID | PMC3408220 |
Grant List | P30-CA014051 / CA / NCI NIH HHS / United States P30-ES002109 / ES / NIEHS NIH HHS / United States R01 CA055042 / CA / NCI NIH HHS / United States R01-CA055042 / CA / NCI NIH HHS / United States U19 ES011399 / ES / NIEHS NIH HHS / United States U54-CA112967 / CA / NCI NIH HHS / United States |