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The reduction of WST-1 reagent on the plasma membrane of epidermal keratinocytes is oxygen-dependent.
MedLine Citation:
PMID:  21354095     Owner:  NLM     Status:  Publisher    
The 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 while 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 cell proliferation assay (which utilizes 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) thus 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 since readings can be affected by certain extrinsic (low 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     Date:  2011-2-23
Journal Detail:
Title:  Analytical biochemistry     Volume:  -     ISSN:  1096-0309     ISO Abbreviation:  -     Publication Date:  2011 Feb 
Date Detail:
Created Date:  2011-2-28     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2011. Published by Elsevier Inc.
Centre for Oncology and Molecular Medicine, Division of Medical Sciences, College of Medicine, Dentistry & Nursing, Ninewells Hospital, University of Dundee, UK.
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