A Multiwell Cardiac μGMEA Platform for Action Potential Recordings from Human iPSC-Derived Cardiomyocyte Constructs

Abstract

SummaryMultielectrode array (MEA) technology has been extensively used for field potential recordings from excitable cells. However, its application for action potential (AP) measurements has not been harnessed. Here, we report a novel platform for high-resolution intracellular AP recordings from induced pluripotent stem cell-cardiomyocyte constructs derived from human cardiac fibroblasts. To gain intracellular access, micro-gold MEAs were used to electroporate multiple constructs simultaneously. High-throughput AP measurements were obtained from 41 multicellular constructs. Repeated electroporations of the same cells did not affect the signal stability. Our model has the capability to distinguish subtle differences in AP morphology to characterize the network profile. Furthermore, we confirm the reliability of the system by recapitulating known drug-induced physiological and arrhythmogenic responses. Overall, the model provides a unique cardio-electronic interface for non-invasive measurements of AP dynamics for drug screening and disease modeling. This technology opens the door for identifying novel cardio-factors to enhance electrophysiological maturation.

Human iPSC-Derived Cardiomyocyte Networks on Multiwell Micro-electrode Arrays for Recurrent Action Potential Recordings

“…It is important to note that batch-to-batch variation in hiPSC differentiation might affect experimental outcomes. The monolayer method of differentiation was optimized in-house for high percent cardiomyocyte production 3,40 . The FACS analysis of MLC2v and TNNT2 markers of our cultures demonstrate a ≥90% ventricular-like phenotype 3 .…”

“…NOTE: This section is intended for thawing and culturing hiPSC-CMs that were differentiated using the feeder-free monolayer method 3,16 and cryopreserved in liquid nitrogen 10 days post-differentiation at 1-2 million cells/vial. Cells from one vial are plated into two substrate-coated wells of a 6-well tissue culture plate.…”

“…Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are the gold standard for an increasing number of laboratories 1,2,3,4,5,6,7,8,9,10 . Beating embryoid bodies 11,12,13 and monolayer 3,7,10,11,12,13,14,15,16,17 differentiation are the preferred methods for cardiomyocyte production and the multielectrode array (MEA) has become a common modality for monitoring the electrodynamics of these networks 18,19,20 . While parameters that can be extracted from field potentials (FPs) such as beating rate, amplitude, duration and RR intervals are baseline electrophysiological responses of spontaneously beating monolayers 18,21,22,23 , the action potential (AP) components underlying these extracellular FP signals are difficult to extrapolate 24 .…”

“…While parameters that can be extracted from field potentials (FPs) such as beating rate, amplitude, duration and RR intervals are baseline electrophysiological responses of spontaneously beating monolayers 18,21,22,23 , the action potential (AP) components underlying these extracellular FP signals are difficult to extrapolate 24 . Our recent publication on the discovery of an application of MEAs for direct recurrent AP measurements provides proof of methodology for exemplary intracellular AP readouts with an extensive waveform analysis at various repolarization phases across multiple batches of hiPSC-derived cardiomyocyte networks 3 . In the study we demonstrated that the delivery of electroporating pulses to networks of hiPSC-derived cardiomyocytes enables intracellular access for AP recordings.…”

“…In the study we demonstrated that the delivery of electroporating pulses to networks of hiPSC-derived cardiomyocytes enables intracellular access for AP recordings. These transient AP recordings are dependent on transmembrane potential recoveries observed through the injury site 3,25,26 . Waveforms recorded via MEA and patch-clamp in our study showed similar AP morphologies thus validating the reliability of the approach 3 .…”