Diermeier-Daucher, Simone and Clarke, Scott T. and Hill, Dani and Vollmann-Zwerenz, Arabel and Bradford, Jolene A. and Brockhoff, Gero (2009) Cell Type Specific Applicability of 5-Ethynyl-2 '-deoxyuridine (EdU) for Dynamic Proliferation Assessment in Flow Cytometry. CYTOMETRY PART A, 75A (6). pp. 535-546. ISSN 1552-4922, 1552-4930
Full text not available from this repository. (Request a copy)Abstract
Using the nucleoside analogue EdU (5-ethynyl-2'-deoxyuridine) for thymidine substitution instead of BrdU (5-bronmo-2'-deoxyuridine) in cell proliferation assays has recently been proposed. However, the effect of EdU on cell viability, DNA synthesis, and cell cycle progression and consequently its usability for dynamic cell proliferation analysis in vitro has not been explored. We compared the effect of EdU and BrdU incorporation into SK-BR-3 and BT474 breast cancer cells and the impact oil cell cycle kinetics, cell viability, and DNA damage. We found that EdU can be used not only for pulse but also for continuous Cell labeling and henceforth in high resolution EdU/Hoechst quenching assays. BrdU and EdU proliferation assays based on click chemistry revealed comparable results. However, cell viability of SK-BR-3 breast cancer cells was highly affected by long term exposure to EdU. Both SK-BR-3 as well as BT474 cells show cell cycle arrests upon long term EdU treatment whereas only SK-BR-3 cells were driven into necrotic cell death by long term exposure to EdU. In contrast BT474 cells appeared essentially unharmed by EdU treatment in terms of viability Consequently using EdU enables highly sensitive and quantitative detection of proliferating cells and facilitates even continuous cell cycle assessment. Nevertheless, potential cellular Susceptibility needs to be individually evaluated. (C) 2009 International Society for Advancement of Cytometry
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | GROWTH-FACTOR RECEPTOR; MAMMALIAN-CELLS; CYCLE ANALYSIS; BREAST-CARCINOMA; CLICK CHEMISTRY; DNA-SYNTHESIS; P53 GENE; BROMODEOXYURIDINE; MUTATIONS; DEOXYCYTIDINE; click chemistry; EdU; BrdU; thymidine analogue; EdU/Hoechst quenching; cell proliferation; BT474; SK-BR-3; cell cycle kinetics; cell death; necrosis |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Frauenheilkunde und Geburtshilfe (Schwerpunkt Frauenheilkunde) Medicine > Lehrstuhl für Pathologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Sep 2020 09:48 |
| Last Modified: | 14 Sep 2020 09:48 |
| URI: | https://pred.uni-regensburg.de/id/eprint/28855 |
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