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

TitleA rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects.
Publication TypeJournal Article
Year of Publication2012
AuthorsValiathan, C, McFaline, JL, Samson, LD
JournalDNA Repair (Amst)
Volume11
Issue1
Pagination92-8
Date Published2012 Jan 2
ISSN1568-7856
KeywordsBiological 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.

DOI10.1016/j.dnarep.2011.11.002
Alternate JournalDNA Repair (Amst.)
PubMed ID22133811
PubMed Central IDPMC3408220
Grant ListP30-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