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Title: Metabolic reduction of resazurin; location within the cell for cytotoxicity assays
Authors: Chen, Jian Lin
Steele, Terry W. J.
Stuckey, David Campbell
Keywords: Science::Biological sciences
Issue Date: 2017
Source: Chen, J. L., Steele, T. W. J., & Stuckey, D. C. (2018). Metabolic reduction of resazurin; location within the cell for cytotoxicity assays. Biotechnology and Bioengineering, 115(2), 351-358. doi:10.1002/bit.26475
Journal: Biotechnology and Bioengineering
Abstract: Resazurin is widely used as a metabolic indicator for living cells, however, there has been considerable debate in the literature with regards to the specific location in the cell where the non-fluorescent resazurin is reduced to the strongly fluorescent resorufin. This lack of clarity about the reduction site makes the use of resazurin reduction data in cytotoxicity studies difficult to interpret. In this study, E. faecalis, a Gram-positive and facultative anaerobic bacterial strain, and the most toxic chlorophenol, pentachlorophenol (PCP), were chosen as models for an anaerobe and toxicant, respectively. By studying the kinetics of resazurin reduction by E. faecalis after different treatments (cell disruption, bacterial filtration, and pre-exposure to toxicant), we confirmed that resazurin reduction to resorufin by live Gram-positive and facultative anaerobic bacterial cells can only happen intracellularly under anaerobic conditions, while resorufin reduction to dihydroresorufin can happen both intracellularly and extracellularly. Based on the understanding of these fundamental mechanisms, we suggest that resazurin reduction can be used as a quick bioassay for measuring cytotoxicity.
ISSN: 0006-3592
DOI: 10.1002/bit.26475
Rights: © 2017 Wiley Periodicals, Inc. All right reserved.
Fulltext Permission: none
Fulltext Availability: No Fulltext
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