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

Abstract

Cardiac safety screening is of paramount importance for drug discovery and therapeutics. Therefore, the development of novel high-throughput electrophysiological approaches for hiPSC-derived cardiomyocyte (hiPSC-CM) preparations is much needed for efficient drug testing. Although multielectrode arrays (MEAs) are frequently employed for field potential measurements of excitable cells, a recent publication by Joshi-Mukherjee and colleagues described and validated its application for recurrent action potential (AP) recordings from the same hiPSC-CM preparation over days. The aim here is to provide detailed step-by-step methods for seeding CMs and for measuring AP waveforms via electroporation with high precision and a temporal resolution of 1 µs. This approach addresses the lack of easy-to-use methodology to gain intracellular access for high-throughput AP measurements for reliable electrophysiological investigations. A detailed work flow and methods for plating of hiPSC-CMs on multiwell MEA plates are discussed emphasizing critical steps wherever relevant. In addition, a custom-built MATLAB script for rapid data handling, extraction and analysis is reported for comprehensive investigation of the waveform analysis to quantify subtle differences in morphology for various AP duration parameters implicated in arrhythmia and cardiotoxicity.

Automated feature extraction from large cardiac electrophysiological data sets

“…High-purity iPSC-CM cultures were produced from human cardiac fibroblasts as described in (Zlochiver et al, 2019). hiPSCs cultures were differentiated using the feederfree monolayer differentiation protocol.…”

“…Much like a medical electrocardiogram, FP recordings measure the potential change across the entire bulk of tissue and have been observed to be closely correlated to the underlying AP (Tertoolen et al, 2018). Recent work has shown the feasibility of multi-electrode arrays (MEA) (Meyer et al, 2012) in the large-scale efficient recording of both AP and FP readings (Edwards et al, 2018;Zlochiver et al, 2019;Hayes et al, 2019;Balafkan et al, 2020).…”

Active Micro-Nano-Collaborative Bioelectronic Device for Advanced Electrophysiological Recording

“…Therefore, several scalable methods for electrophysiological signal recording have been explored, including passive electrodes [31][32][33][34][35][36][37] and active micro/nanobioelectronic devices [38][39][40][41][42][43][44][45][46]. Microelectrode arrays (MEAs) [31,[47][48][49][50][51][52] are passive electrodes that are commonly produced on insulating substrates. These electrodes receive electrical signals from cells and subsequently transmit them to external amplifiers through leads that are coated with passivation layers.…”

Integrated Cardiomyocyte-Based Biosensing Platform for Electroporation-Triggered Intracellular Recording in Parallel with Delivery Efficiency Evaluation

“…Cardiomyocytes are participating in the regulation of the contraction–relaxation period of the heart. Their self-regulation of contractility and the generation of action potential involve transportation by multiple ion channels and exchangers, which are of high sensitivity to external electrical stimulation. − Microelectrode array (MEA)-based extracellular recording featured noninvasive, dynamic, long-term, and large-scale electrophysiological studies, but it does not provide important information such as resting membrane potential and ion channel conductance. − Electroporation is a well-established technique that induces the generation of nanopores in the cell membrane for intracellular action potential recording access. − Due to the accessibility and high throughput, electroporation is also one of the most common physical techniques for biomolecule delivery (e.g., mRNA, DNA, and proteins) which holds great potential for genetic engineering, cellular imaging, and medical clinical applications. − Compared to virus-mediated and chemical methods, − electroporation prevents toxicity to cells . However, bulk electroporation (BEP)-based delivery of cardiomyocytes remains challenging.…”