title: A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes.
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.
authors.affiliation: Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah School of Medicine, Salt Lake City, Utah;
authors.forename: Angelica
authors.initials: A
authors.lastname: Lopez-Izquierdo
authors.affiliation: Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah School of Medicine, Salt Lake City, Utah;
authors.forename: Mark
authors.initials: M
authors.lastname: Warren
authors.affiliation: Cardiovascular Center, Medical College of Wisconsin, Milwaukee, Wisconsin; and.
authors.forename: Michael
authors.initials: M
authors.lastname: Riedel
authors.affiliation: Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah School of Medicine, Salt Lake City, Utah;
authors.forename: Scott
authors.initials: S
authors.lastname: Cho
authors.affiliation: Cardiovascular Center, Medical College of Wisconsin, Milwaukee, Wisconsin; and.
authors.forename: Shuping
authors.initials: S
authors.lastname: Lai
authors.affiliation: Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah School of Medicine, Salt Lake City, Utah;
authors.forename: Robert L
authors.initials: RL
authors.lastname: Lux
authors.affiliation: Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah School of Medicine, Salt Lake City, Utah;
authors.forename: Kenneth W
authors.initials: KW
authors.lastname: Spitzer
authors.affiliation: Cardiovascular Center, Medical College of Wisconsin, Milwaukee, Wisconsin; and.
authors.forename: Ivor J
authors.initials: IJ
authors.lastname: Benjamin
authors.affiliation: Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah School of Medicine, Salt Lake City, Utah; Division of Pediatric Cardiology, University of Utah School of Medicine, Salt Lake City, Utah mfirouzi@cvrti.utah.edu.
authors.forename: Martin
authors.initials: M
authors.lastname: Tristani-Firouzi
authors.affiliation: Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah School of Medicine, Salt Lake City, Utah; Division of Pediatric Cardiology, University of Utah School of Medicine, Salt Lake City, Utah.
authors.forename: Chuanchau J
authors.initials: CJ
authors.lastname: Jou
mesh_terms: D015081:2-Naphthylamine / Q000031:analogs & derivatives* / Q000737:chemistry; D000200:Action Potentials; D002454:Cell Differentiation; D005456:Fluorescent Dyes; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology*; D007259:Infrared Rays; D032383:Myocytes, Cardiac / Q000166:cytology / Q000502:physiology*; D011806:Quinolinium Compounds / Q000737:chemistry*; D056969:Voltage-Sensitive Dye Imaging / Q000379:methods*
chemical_list: C553754:1-(4-sulfanatobutyl)-4-(beta-(2-(di-n-butylamino)-6-naphthyl)butadienyl)quinolinium betaine; D005456:Fluorescent Dyes; D011806:Quinolinium Compounds; D015081:2-Naphthylamine
keywords: QT prolongation; acquired long QT syndrome; optical action potential; preclinical drug screening; stem cell model