Combining electrophysiology and contractility recordings for more complete assessment of hiPSC-CMs

Europhysiology 2018 (London, UK) (2018) Proc Physiol Soc 41, PCB065

Poster Communications: Combining electrophysiology and contractility recordings for more complete assessment of hiPSC-CMs

A. R. Obergrussberger1, S. Stoelzle-Feix1, E. Dragicevic1, N. Becker1, K. Juhasz1, U. Thomas1, L. Doerr1, M. Rapedius1, C. Haarmann1, I. Rinke-Weiß1, T. A. Goetze1, M. Beckler1, M. George1, A. Brüggemann1, N. Fertig1

1. Nanion Technologies GmbH, Munich, Germany.

View other abstracts by:


Human induced pluripotent stem cells (hiPSCs) are becoming increasingly important for cardiac safety testing due to their recapitulation of native behaviour, relative abundance and ease-of-use. We combine automated patch clamp (APC), impedance and extracellular field potential (EFP) measurements to study hiPSC-derived cardiomyocytes (hiPSC-CMs) from different sources. In line with the comprehensive in vitro proarrhythmia assay (CiPA) initiative, cardiac ion channels expressed in heterologous expression systems have been recorded on APC devices at room temperature and at physiological temperature and the effects of different pro-arrhythmic compounds on ion channels was investigated. In addition hiPSC-CMs were used on APC platforms recording from 1, 8 or 384 wells simultaneously. Voltage-gated Na+ (NaV), Ca2+ (CaV) and K+ (KV) currents were recorded. The NaV channel was blocked by tetracaine with an IC50 of 2.9 ± 1.4 µM (n = 5) and by TTX with an IC50 of 1.1 ± 0.5 µM (n = 4), values which are consistent with the cardiac NaV channel, NaV1.5. Nifedipine blocked the CaV currents recorded from hiPSC-CMs with an IC50 of 84 nM, consistent with the cardiac CaV1.2 channel. In addition to voltage clamp measurements, action potentials from hiPSC-CMs were also recorded in the current clamp mode and the effect of nifedipine on action potential amplitude and duration will be shown. Using the dynamic clamp technique coupled with an APC device, electrically modelled IK1 was injected into the cell under current clamp conditions and this resulted in a more hyperpolarized and stable resting membrane potential. The amount of IK1 injected into the cell affected action potential (AP) duration. Pharmacology using a calcium channel activator, BayK 8644, and blocker, nifedipine, was also performed. BayK 8644 at 1 µM significantly prolonged AP duration and nifedipine (30 µM) shortened AP duration. Within the myocyte phase II study of the CiPA initiative, a device combining impedance-based contractility and extracellular field potential (EFP) recordings was used to investigate the effects of different compounds deemed low, intermediate and high risk by the FDA. Different hiPSC-CMs were used in a large cross-site evaluation which found that high risk compounds such as dofetilide prolong field potential duration and arrhythmic events were detected in both impedance and EFP recordings. Intermediate risk compounds induced arrhythmia in almost all cases at the highest dose and either no significant effect or a decrease in FPDmax was recorded for low risk compounds. In addition, cells can be optically stimulated and impedance and EFP signals simultaneously recorded from the same cellular monolayer. In this way, contractility and EFP signals can be exactly overlaid giving in-depth mechanistic information about contractile behaviour and electrophysiology of iPSC-CMs.



Where applicable, experiments conform with Society ethical requirements.

Site search

Filter

Content Type