Document Detail

The reduction of water-soluble tetrazolium salt reagent on the plasma membrane of epidermal keratinocytes is oxygen dependent.
MedLine Citation:
PMID:  21354095     Owner:  NLM     Status:  MEDLINE    
The water-soluble tetrazolium salt (WST-1) assay is frequently used to assess cell proliferation. However, our study showed that in normal and cancerous keratinocytes, this assay is more responsive to changes in oxygenation than to rates of cell growth. Stimulation of keratinocyte proliferation by low Ca(2+) and suppression of proliferation by nocodazole resulted in modest changes in WST-1 readings, whereas gradually reducing the level of oxygen in the cellular environment from ambient (21%) to near anoxic (0.1%) revealed a very strong negative correlation between cell oxygenation and WST-1 reagent reduction. In contrast, the very similar MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cell proliferation assay, which uses a different tetrazolium salt, showed no sensitivity to the level of oxygen. Unlike MTT, WST-1 reagent is reduced extracellularly through trans-plasma membrane transport (tPMET), thereby suggesting that tPMET is oxygen dependent. We propose that the WST-1 assay can be developed into a sensitive quantitative method to evaluate cell oxygenation in vitro and used to study the role of hypoxia and tPMET in homeostasis and disease (e.g., cancer). At the same time, WST-1 assay should be used cautiously to assess cell viability or proliferation because readings can be affected by certain extrinsic (low atmospheric oxygen or high density culture) or intrinsic (defects in oxygen-sensing pathways) factors.
Lynda Weir; Douglas Robertson; Irene M Leigh; Andrey A Panteleyev
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2011-02-24
Journal Detail:
Title:  Analytical biochemistry     Volume:  414     ISSN:  1096-0309     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2011 Jul 
Date Detail:
Created Date:  2011-05-09     Completed Date:  2011-08-22     Revised Date:  2014-07-22    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  31-7     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 Elsevier Inc. All rights reserved.
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MeSH Terms
Cell Hypoxia
Cell Line, Tumor
Cell Membrane / metabolism*
Cell Proliferation* / drug effects
Cells, Cultured
Indicators and Reagents / metabolism
Keratinocytes / cytology*,  metabolism
Nocodazole / pharmacology
Oxygen / metabolism*
Tetrazolium Salts / metabolism*
Tubulin Modulators / pharmacology
Grant Support
13044//Cancer Research UK; //Cancer Research UK
Reg. No./Substance:
0/2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; 0/Indicators and Reagents; 0/Tetrazolium Salts; 0/Tubulin Modulators; S88TT14065/Oxygen; SH1WY3R615/Nocodazole

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