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- W2912686758 abstract "Accurate measurement of the effect of drugs on the action potential (AP) is critical for drug safety screening and research. Cellular leak currents increase during voltage-clamp recordings, and can lead to instability and errors. We determined the effect of leak current on resting membrane potential (RMP) and AP characteristics, as well as how changes in leak affects observed APs during drug-cell interactions. We recorded APs from human induced pluripotent stem cell-derived cardiac myocytes (hiPSC-CMs) and used the Cybercyte dynamic clamp system to electronically express IK1, which gives hiPSC-CMs physiological RMP and enhanced AP stability. Additionally, we used the Cybercyte to express real-time electronic leak current correction (ILCC). As leak current increases, RMP depolarizes, AP morphology changes and, eventually, APs cannot be evoked. Electronic expression of leak current correction, ILCC, restored RMP to ∼-84 mV and resulted in stable APs. Dofetilide, which blocks IKr and increases AP duration (APD), and nifedipine, which blocks ICa, and decreases APD, were applied to cells with and without leak current correction. 30nM dofetilide prolonged APD, but the effect was exaggerated with leak current (51±25%, n=6), compared with cells with leak current correction, (ILCC: 34±12%, n=8). Conversely, 100 nM nifedipine shortened APD, but the effect was blunted with cell leak (11±28%, n=4), compared with leak current corrected cells (+ILCC: 45±9%, n=5). Leak current correction also significantly reduced APD measurement variability. Our data demonstrate that real-time leak current correction combined with electronic addition of IK1 improves the accuracy of measuring the effects of dofetilide and nifedipine on AP morphology and improves the efficiency by reducing measurement variability and the number of data points required to determine the effect of drugs on the AP." @default.
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- W2912686758 date "2019-02-01" @default.
- W2912686758 modified "2023-10-17" @default.
- W2912686758 title "Effect of Real-Time Leak Current Correction on Action Potentials Recorded from Induced Pluripotent Stem Cell-Derived Cardiac Myocytes" @default.
- W2912686758 doi "https://doi.org/10.1016/j.bpj.2018.11.577" @default.
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