Abstract
Standard acute cardiac safety pharmacological assays usually comprise high-throughput screening (HTS) of single cardiac ion channels like the hERG channel expressed in tumor cell lines and low-throughput assay systems like ex vivo tissue preparations or Langendorff perfused hearts from animals. These types of model systems typically lack sufficient predictivity and/or throughput.
In contrast, standardized and pure human induced pluripotent stem cell-derived cardiomyocytes (hiPSCM) are commercially available and have already been demonstrated to be predictive human cell models in which to assess cardiotoxicity in vitro (Obejero-Paz et al., Sci Rep 5:17623, 2015; Lu et al., Toxicol Sci 148:503–516, 2015; Kitaguchi et al., J Pharmacol Toxicol Methods 78:93–102, 2015; Gilchrist et al., Toxicol Appl Pharmacol 288:249–257, 2015).
Here it is described how human-induced pluripotent stem cell-derived Cor.4U cardiomyocytes can be implemented in an HTS assay environment using the FDSS plate reader platforms from Hamamatsu and modern fluorescent probes to monitor drug-induced changes to either cardiac cytosolic free calcium ([Ca2+]i) transients or plasma membrane potentials.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Obejero-Paz CA, Bruening-Wright A et al (2015) Quantitative profiling of the effects of vanoxerine on human cardiac ion channels and its application to cardiac risk. Sci Rep 5:17623. doi:10.1038/srep17623
Lu HR, Whittaker R et al (2015) High throughput measurement of Ca++ dynamics in human stem cell-derived cardiomyocytes by kinetic image cytometery: a cardiac risk assessment characterization using a large panel of cardioactive and inactive compounds. Toxicol Sci 148(2):503–516. doi:10.1093/toxsci/kfv201
Kitaguchi T, Moriyama Y et al (2015) CSAHi study: evaluation of multi-electrode array in combination with human iPS cell-derived cardiomyocytes to predict drug-induced QT prolongation and arrhythmia—effects of 7 reference compounds at 10 facilities. J Pharmacol Toxicol Methods 78:93–102. doi:10.1016/j.vascn.2015.12.002
Gilchrist KH, Lewis GF et al (2015) High-throughput cardiac safety evaluation and multi-parameter arrhythmia profiling of cardiomyocytes using microelectrode arrays. Toxicol Appl Pharmacol 288(2):249–257. doi:10.1016/j.taap.2015.07.024
Fermini B, Hancox JC et al (2016) A new perspective in the field of cardiac safety testing through the comprehensive in vitro proarrhythmia assay paradigm. J Biomol Screen 21(1):1–11. doi:10.1177/1087057115594589
Maher MP, Gonzales JE (2005) Multi well plate and microelectrode assemblies for ion channel assays. US Patent 6,969,449B2, 29 Nov 2005
Lundholt BK, Scudder KM, Pagliaro L (2003) A simple technique for reducing edge effect in cell-based assays. J Biomol Screen 8(5):566–570. doi:10.1177/1087057103256465
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kettenhofen, R. (2017). HTS-Compatible Voltage- and Ca2+-Sensitive Dye Recordings from hiPSC-Derived Cardiomyocytes Using the Hamamatsu FDSS Systems. In: Clements, M., Roquemore, L. (eds) Stem Cell-Derived Models in Toxicology. Methods in Pharmacology and Toxicology. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6661-5_7
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6661-5_7
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6659-2
Online ISBN: 978-1-4939-6661-5
eBook Packages: Springer Protocols