期刊
AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY
卷 307, 期 9, 页码 H1370-H1377出版社
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/ajpheart.00344.2014
关键词
optical action potential; preclinical drug screening; QT prolongation; acquired long QT syndrome; stem cell model
资金
- Nora Eccles Treadwell Foundation
- American Heart Association
- National Heart, Lung, and Blood Institute (NHLBI) National Institutes of Health Director's Pioneer Award [7-DP1-HL-117650-05]
- Veterans Affairs Merit Review Award
- NHLBI Grant [5-R01-HL-074370-04, R37-HL-042873-24]
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-{beta[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.
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