1. A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes
- Author
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Robert L. Lux, Chuanchau J. Jou, Scott Cho, Mark Warren, Ivor J. Benjamin, Angelica Lopez-Izquierdo, Shuping Lai, Michael Riedel, Kenneth W. Spitzer, and Martin Tristani-Firouzi
- Subjects
Physiology ,Infrared Rays ,Cellular differentiation ,Quinolinium Compounds ,Induced Pluripotent Stem Cells ,Voltage-sensitive dye ,Action Potentials ,Cell Differentiation ,Pharmacology ,Biology ,Fluorescence ,QT interval ,Voltage-Sensitive Dye Imaging ,Electrophysiology ,Integrative Cardiovascular Physiology and Pathophysiology ,Physiology (medical) ,2-Naphthylamine ,Biophysics ,Repolarization ,Humans ,Myocytes, Cardiac ,Cardiology and Cardiovascular Medicine ,Induced pluripotent stem cell ,Fluorescent Dyes - 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.
- Published
- 2014