Information de reference pour ce titreAccession Number: | 00115742-201612000-00013.
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Author: | Hortigon-Vinagre, M. P.; Zamora, V.; Burton, F. L.; Green, J.; Gintant, G. A.; Smith, G. L.
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Institution: | (*)Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Science, University of Glasgow 126 University Place, Glasgow G12 8TA, United Kingdom (+)Clyde Biosciences Ltd, BioCity Scotland, Bo'Ness Road, Newhouse, Lanarkshire, Scotland ML1 5UH, United Kingdom (++)AbbVie, 1 North Waukegan Road, Department ZR-13, Building AP-9A, North Chicago, Illinois 60064-6119
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Title: | The Use of Ratiometric Fluorescence Measurements of the Voltage Sensitive Dye Di-4-ANEPPS to Examine Action Potential Characteristics and Drug Effects on Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.[Miscellaneous Article]
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Source: | Toxicological Sciences. 154(2):320-331, December 2016.
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Abstract: | Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) and higher throughput platforms have emerged as potential tools to advance cardiac drug safety screening. This study evaluated the use of high bandwidth photometry applied to voltage-sensitive fluorescent dyes (VSDs) to assess drug-induced changes in action potential characteristics of spontaneously active hiPSC-CM. Human iPSC-CM from 2 commercial sources (Cor.4U and iCell Cardiomyocytes) were stained with the VSD di-4-ANEPPS and placed in a specialized photometry system that simultaneously monitors 2 wavebands of emitted fluorescence, allowing ratiometric measurement of membrane voltage. Signals were acquired at 10 kHz and analyzed using custom software. Action potential duration (APD) values were normally distributed in cardiomyocytes (CMC) from both sources though the mean and variance differed significantly (APD90: 229 +/- 15 ms vs 427 +/- 49 ms [mean +/- SD, P < 0.01]; average spontaneous cycle length: 0.99 +/- 0.02 s vs 1.47 +/- 0.35 s [mean +/- SD, P < 0.01], Cor.4U vs iCell CMC, respectively). The 10-90% rise time of the AP (Trise) was ~6 ms and was normally distributed when expressed as 1/ in both cell preparations. Both cell types showed a rate dependence analogous to that of adult human cardiac cells. Furthermore, nifedipine, ranolazine, and E4031 had similar effects on cardiomyocyte electrophysiology in both cell types. However, ranolazine and E4031 induced early after depolarization-like events and high intrinsic firing rates at lower concentrations in iCell CMC. These data show that VSDs provide a minimally invasive, quantitative, and accurate method to assess hiPSC-CM electrophysiology and detect subtle drug-induced effects for drug safety screening while highlighting a need to standardize experimental protocols across preparations.
(C) Society of Toxicology 2016. Published by Oxford University Press. All rights reserved.
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Author Keywords: | methods; human induced pluripotent stem cell-derived cardiomyocytes; stem cells; action potential duration; voltage sensitive dye; drug screening.
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Language: | English.
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Document Type: | Voltage-Sensitive Dyes in Human iPSC-Derived Cardiomyocytes.
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Journal Subset: | Clinical Medicine.
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ISSN: | 1096-6080
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NLM Journal Code: | cz1, 9805461
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DOI Number: | https://dx.doi.org/10.1093/toxsc...- ouverture dans une nouvelle fenêtre
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