High-Throughput Phenotyping of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes and Neurons Using Electric Field Stimulation and High-Speed Fluorescence Imaging

Assay and Drug Development Technologies
Neil J DailyTetsuro Wakatsuki

Abstract

Electrophysiology of excitable cells, including muscle cells and neurons, has been measured by making direct contact with a single cell using a micropipette electrode. To increase the assay throughput, optical devices such as microscopes and microplate readers have been used to analyze electrophysiology of multiple cells. We have established a high-throughput (HTP) analysis of action potentials (APs) in highly enriched motor neurons and cardiomyocytes (CMs) that are differentiated from human induced pluripotent stem cells (iPSCs). A multichannel electric field stimulation (EFS) device enabled the ability to electrically stimulate cells and measure dynamic changes in APs of excitable cells ultra-rapidly (>100 data points per second) by imaging entire 96-well plates. We found that the activities of both neurons and CMs and their response to EFS and chemicals are readily discerned by our fluorescence imaging-based HTP phenotyping assay. The latest generation of calcium (Ca2+) indicator dyes, FLIPR Calcium 6 and Cal-520, with the HTP device enables physiological analysis of human iPSC-derived samples highlighting its potential application for understanding disease mechanisms and discovering new therapeutic treatments.

References

Aug 1, 1992·Journal of the American College of Cardiology·B Surawicz, C Fisch
Oct 1, 1990·Pacing and Clinical Electrophysiology : PACE·K M KavanaghD G Wyse
Oct 26, 1999·Cardiovascular Research·D E Euler
Feb 13, 2001·Current Opinion in Biotechnology·D L TaylorK A Giuliano
Apr 6, 2002·Circulation Research·Vladimir G Fast, Eric R Cheek
Sep 1, 1951·The Journal of Physiology·M H DRAPER, S WEIDMANN
May 25, 2004·Circulation Research·Vladimir G FastRaymond E Ideker
Jul 10, 2004·Circulation Research·Igor R EfimovGuy Salama
Jan 26, 2005·Trends in Neurosciences·Shigetada Nakanishi
Jan 19, 2006·Biochemical Society Transactions·H BetzV Eulenburg
Dec 26, 2006·Expert Review of Cardiovascular Therapy·Henri Roukoz, Walid Saliba
Sep 12, 2008·Tissue Engineering. Part a·Loraine L Y ChiuMilica Radisic
Dec 23, 2008·Drug Discovery Today·James L Stevens, Thomas K Baker
Feb 7, 2012·Proceedings of the National Academy of Sciences of the United States of America·Evan W MillerRoger Y Tsien
Feb 24, 2012·Clinical Neurophysiology : Official Journal of the International Federation of Clinical Neurophysiology·C J Stam, E C W van Straaten
Apr 12, 2013·Proceedings of the Japan Academy. Series B, Physical and Biological Sciences·Kunihiko Obata
Aug 2, 2013·Nature Reviews. Drug Discovery·Kelly Rae Chi
Dec 11, 2013·American Journal of Physiology. Heart and Circulatory Physiology·Faisal M MerchantAntonis A Armoundas
Jul 22, 2014·Clinical and Experimental Pharmacology & Physiology·Joshua N Edwards, Lothar A Blatter
Apr 15, 2015·Proceedings of the National Academy of Sciences of the United States of America·Cedric BardyFred H Gage
Jul 21, 2015·Scientific Reports·Huisheng LiuSu-Chun Zhang
Aug 8, 2015·Journal of the American Chemical Society·Vincent GrenierEvan W Miller
Dec 15, 2015·Acta Pharmacologica Sinica·Hai-bo YuXiao-liang Wang
Oct 3, 2017·Journal of Evolving Stem Cell Research·Neil J DailyTetsuro Wakatsuki

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Citations

Oct 5, 2018·Frontiers in Cardiovascular Medicine·Michael J GreenbergTetsuro Wakatsuki
Oct 30, 2021·Frontiers in Genome Editing·Amy McTagueManju A Kurian

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Methods Mentioned

BETA
fluorescence

Software Mentioned

MATLAB
InvivoSciences

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