A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes

作者：Lopez Izquierdo Angelica; Warren Mark; Riedel Michael; Cho Scott; Lai Shuping; Lux Robert L; Spitzer Kenneth W; Benjamin Ivor J; Tristani Firouzi Martin^*; Jou Chuanchau J

来源：American Journal of Physiology - Heart and Circulatory Physiology, 2014, 307(9): H1370-H1377.

DOI：10.1152/ajpheart.00344.2014

• optical action potential
• preclinical drug screening
• QT prolongation
• acquired long QT syndrome
• stem cell model

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 %26gt; 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.

• 出版日期2014-11-1