A rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects
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 t...
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| Format: | Article |
| Language: | en_US |
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Elsevier
2015
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| Online Access: | http://hdl.handle.net/1721.1/99508 https://orcid.org/0000-0002-7112-1454 |
| _version_ | 1810975839560400896 |
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| author | Valiathan, Chandni McFaline, Jose Luis Samson, Leona D. |
| author2 | Massachusetts Institute of Technology. Center for Environmental Health Sciences |
| author_facet | Massachusetts Institute of Technology. Center for Environmental Health Sciences Valiathan, Chandni McFaline, Jose Luis Samson, Leona D. |
| author_sort | Valiathan, Chandni |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T08:45:45Z |
| format | Article |
| id | mit-1721.1/99508 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:45:45Z |
| publishDate | 2015 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/995082022-09-23T14:22:50Z A rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects Valiathan, Chandni McFaline, Jose Luis Samson, Leona D. 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 Valiathan, Chandni McFaline, Jose Luis Samson, Leona D. 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. National Institutes of Health (U.S.) (U54-CA112967) National Institutes of Health (U.S.) (R01-CA055042) National Institutes of Health (U.S.) (P30-ES002109) National Institutes of Health (U.S.) (P30-CA014051) Massachusetts Institute of Technology (Merck Fellowship) 2015-10-29T17:12:29Z 2015-10-29T17:12:29Z 2011-11 2011-10 Article http://purl.org/eprint/type/JournalArticle 15687864 http://hdl.handle.net/1721.1/99508 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. https://orcid.org/0000-0002-7112-1454 en_US http://dx.doi.org/10.1016/j.dnarep.2011.11.002 DNA Repair Creative Commons Attribution http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Valiathan, Chandni McFaline, Jose Luis Samson, Leona D. A rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects |
| title | A rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects |
| title_full | A rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects |
| title_fullStr | A rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects |
| title_full_unstemmed | A rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects |
| title_short | A rapid survival assay to measure drug-induced cytotoxicity and cell cycle effects |
| title_sort | rapid survival assay to measure drug induced cytotoxicity and cell cycle effects |
| url | http://hdl.handle.net/1721.1/99508 https://orcid.org/0000-0002-7112-1454 |
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