A New Fluorometric Assay for Cytotoxicity Measurements in-Vitro

Page, Benedict J.; Page, Melissa M.; Noël, Christiane

doi:10.3892/ijo.3.3.473

Cited by 328 publications

(343 citation statements)

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“…At 7 days PI the cells on cover slips were fixed with 4% paraformaldehyde/0.25% glutaraldehyde and ␤-galactosidase detected by exposure to 1% X-gal reagent solution for 5-6 h. Cultured cells were exposed to Alamar blue (Alamar Biosciences, Westlake, OH, USA) (40 l/ml) overnight at 4°C and cell survival determined by quantitative fluorescence (530/590 nm excitation/emission). 9 For RNA analysis, the neurons were lysed in TRI reagent (Sigma Chemical), and RNA resuspended in 20 l distilled water. For RT-PCR reactions 5-10 l of each sample was used.…”

Section: Construction Of Complementing Cell Lines and Viral Mutantsmentioning

confidence: 99%

Deletion of multiple immediate–early genes from herpes simplex virus reduces cytotoxicity and permits long-term gene expression in neurons

et al. 1998

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Section: Construction Of Complementing Cell Lines and Viral Mutantsmentioning

confidence: 99%

Deletion of multiple immediate–early genes from herpes simplex virus reduces cytotoxicity and permits long-term gene expression in neurons

et al. 1998

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Section: A Brief Recap Of Cytotoxicity Assaysmentioning

confidence: 99%

“…Other tetrazolium salts that yield a soluble formazan followed, along with the introduction of intermediary electron acceptors, such as phenazine methosulfate, to facilitate dye reduction [8]. The sensitivity of dye approaches was increased with a halving of assay time and reduction of costs by introduction of the Alamar Blue assay [9], which had a good correlation with neutral red uptake, LDH release, total protein, and cell density [10].Other viability assays were developed that monitored changes in intracellular activity. Rhodamine-123 was the first mitochondrial membrane potential dye to become available and provided an earlier indicator of the loss of cell viability compared with trypan blue [11].…”

mentioning

confidence: 99%

High-content analysis for drug delivery and nanoparticle applications

Brayden

Cryan

Dawson

et al. 2015

Drug Discovery Today

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Publication information Drug Discovery Today, 20 (8): 942-957Publisher Elsevier Item record/more information http://hdl.handle.net/10197/6636 Publisher's statementThis is the author's version of a work that was accepted for publication in Drug Discovery Today. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Drug Discovery Today, 20 (8), (2015 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Page 1 of 26A c c e p t e d M a n u s c r i p t Sally-Ann CryanSally-Ann Cryan has a pharmacy degree and a PhD in pharmaceutics ( Jeremy SimpsonJeremy Simpson carried out his PhD at the University of Warwick, followed postdoctoral work at the Scripps Research Institute in San Diego, and the Imperial Cancer Research Fund in London. After 9 years as a staff member at the European Molecular Biology Laboratory (EMBL) in Heidelberg (Germany), he was appointed as professor of cell biology at UCD, in 2008. He currently applies high-throughput imaging technologies to study subcellular transport pathways and the internalization routes taken by nanoparticles in cells. He runs the UCD Cell Screening Laboratory and is the author of more than 80 publications.High-content analysis (HCA) provides quantitative multiparametric cellular fluorescence data. From its origins in discovery toxicology, it is now addressing fundamental questions in drug delivery. Nanoparticles (NPs), polymers, and intestinal permeation enhancers are being harnessed in drug delivery systems to modulate plasma membrane properties and the intracellular environment. Identifying comparative mechanistic cytotoxicity on sublethal events is crucial to expedite the development of such systems. NP uptake and intracellular routing pathways are also being dissected using chemical and genetic perturbations, with the potential to assess the intracellular fate of targeted and untargeted particles in vitro. As we discuss here, HCA is set to make a major impact in preclinical delivery research by elucidating the intracellular pathways of NPs and the in vitro mechanistic-based toxicology of formulation constituents. A brief recap of cytotoxicity assaysIn vitro cell-based assays have long been used to assess the cytotoxic effects of drug exposure [1]. Cells undergoing acute necrosis swell and lose metabolic capacity, thereby losing the capacity to maintain a barrier to the extracellular space and the ability ...

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“…The AlamarBlue ® assay (ThermoScientific, Waltham, MA USA) is a reliable method for measuring cell viability (6). This assay, using the metabolic activity of cells to reduce resazurin (oxidized form: 7-hydroxy-3H-phenoxazin-3-1-10-oxide) to resorufin, counts the number of cells with active mitochondria, as resazurin reduction is achieved by mitochondrial enzymes (7). The fluorescence of these two forms is measured at 530-560nm as excitation wavelength and at 590nm emission wavelength.…”

Section: Methodsmentioning

confidence: 99%