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A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes. Am J Physiol Heart Circ Physiol 2014 Nov 01;307(9):H1370-7

Date

08/31/2014

Pubmed ID

25172899

Pubmed Central ID

PMC4217013

DOI

10.1152/ajpheart.00344.2014

Scopus ID

2-s2.0-84908375059   23 Citations

Abstract

Human induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM)-based assays are emerging as a promising tool for the in vitro preclinical screening of QT interval-prolonging side effects of drugs in development. A major impediment to the widespread use of human iPSC-CM assays is the low throughput of the currently available electrophysiological tools. To test the precision and applicability of the near-infrared fluorescent voltage-sensitive dye 1-(4-sulfanatobutyl)-4-{β[2-(di-n-butylamino)-6-naphthyl]butadienyl}quinolinium betaine (di-4-ANBDQBS) for moderate-throughput electrophysiological analyses, we compared simultaneous transmembrane voltage and optical action potential (AP) recordings in human iPSC-CM loaded with di-4-ANBDQBS. Optical AP recordings tracked transmembrane voltage with high precision, generating nearly identical values for AP duration (AP durations at 10%, 50%, and 90% repolarization). Human iPSC-CMs tolerated repeated laser exposure, with stable optical AP parameters recorded over a 30-min study period. Optical AP recordings appropriately tracked changes in repolarization induced by pharmacological manipulation. Finally, di-4-ANBDQBS allowed for moderate-throughput analyses, increasing throughput >10-fold over the traditional patch-clamp technique. We conclude that the voltage-sensitive dye di-4-ANBDQBS allows for high-precision optical AP measurements that markedly increase the throughput for electrophysiological characterization of human iPSC-CMs.

Author List

Lopez-Izquierdo A, Warren M, Riedel M, Cho S, Lai S, Lux RL, Spitzer KW, Benjamin IJ, Tristani-Firouzi M, Jou CJ

Author

Ivor J. Benjamin MD Center Director, Professor in the Medicine department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

2-Naphthylamine
Action Potentials
Cell Differentiation
Fluorescent Dyes
Humans
Induced Pluripotent Stem Cells
Infrared Rays
Myocytes, Cardiac
Quinolinium Compounds
Voltage-Sensitive Dye Imaging
jenkins-FCD Prod-482 91ad8a360b6da540234915ea01ff80e38bfdb40a