Document Detail

Use of the Viability Reagent PrestoBlue in Comparison with Alamarblue and MTT to Assess the Viability of Human Corneal Epithelial Cells.
MedLine Citation:
PMID:  25464019     Owner:  NLM     Status:  Publisher    
INTRODUCTION: PrestoBlue is a new resazurin based reagent to assess cell viability and cytotoxicity. It is claimed to be a fast and highly sensitive assay. Here, we compared PrestoBlue, alamarBlue, and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazonium bromide (MTT) in assessing cell viability of human corneal epithelial cells (HCEC), and investigated the effect of plate color, reading mode, and plate storage on the performance of PrestoBlue assay.
METHODS: The viability of different numbers of healthy HCEC and the toxicity of various chemicals on HCEC were evaluated using PrestoBlue (fluorescence), alamarBlue(fluorescence), and MTT (absorbance). The sensitivities of the three assays were compared. In the PrestoBlue assay, three plate colors and two reading modes were used and compared in assessing the toxic effect of SDS. The PrestoBlue solutions after reaction were stored and measured on day 1, 2, 3, 5, and 7. The fluorescence readings obtained on different days were then compared.
RESULTS: Both PrestoBlue and alamarBlue were able to detect 5000 healthy cells after 30min incubation and 1000 cells after 1h, 2h, and 4h incubation; while MTT was able to detect 5000 cells after 3h incubation. In the assessment of the toxicity of various chemicals, PrestoBlue and alamarBlue performed similarly. There was no significant difference between the results obtained by these two reagents. All the three plate colors and two reading modes showed similar results in the PrestoBlue assay in assessing the toxicity of SDS. Plate storage up to 7days did not affect the result of the PrestoBlue assay.
CONCLUSION: Our study suggests that in evaluating the viability of HCEC, PrestoBlue is more sensitive than MTT, but similar to alamarBlue. The plate color, reading mode and plate storage up to 7days did not affect the performance of the PrestoBlue assay.
Manlong Xu; David J McCanna; Jacob G Sivak
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-11-15
Journal Detail:
Title:  Journal of pharmacological and toxicological methods     Volume:  71C     ISSN:  1873-488X     ISO Abbreviation:  J Pharmacol Toxicol Methods     Publication Date:  2014 Nov 
Date Detail:
Created Date:  2014-12-2     Completed Date:  -     Revised Date:  2014-12-3    
Medline Journal Info:
Nlm Unique ID:  9206091     Medline TA:  J Pharmacol Toxicol Methods     Country:  -    
Other Details:
Languages:  ENG     Pagination:  1-7     Citation Subset:  -    
Copyright Information:
Copyright © 2014. Published by Elsevier Inc.
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