Automated Patch Clamp on mESC-Derived Cardiomyocytes for Cardiotoxicity Prediction

Abstract

IntroductIon Studying cardiac action potential modulation is important for preclinical drug safety testing. it offers a rationale for potentially required medicinal chemistry efforts, lead compound optimization, or project termination due to cardiotoxic side effects.the patch clamp technique is the gold standard for real-time investigation of ion channels, 1 and the automation of the method increases the throughput and makes it accessible to a wider audience due to its ease of use compared with the conventional patch clamp method. 2 to overcome the hurdles in working with primary heart tissue from human donors or animals, the use of stem cell-derived cardiomyocytes may be a viable option for safety screening of larger-compound libraries in the earlier stages of the drug development process.Here we describe the electrophysiological characterization of mouse embryonic stem cell (meSc)-derived cardiomyocytes on an automated patch clamp platform with voltage clamp and current clamp capability for assessing action potentials and their pharmacological modulation.these cardiomyocytes are readily available and 100% pure, in contrast to preparations of primary cardiac myocytes, which are usually contaminated by fibroblasts. 3 moreover, the selected meSc-derived cardiomyocytes revealed their physiological relevance by functional integration into in farcted mouse hearts and prolonging the span of life of transplanted mice in comparison with the control group. 4 We show that these meSc-derived cardiomyocytes functionally express all essential cardiac ion channels and that typical cardiac action potentials can be elicited.the combination of an automated patch clamp instrument together with a standardized and pure cardiac myocyte model enables scientists in basic or applied cardiology and toxicology to perform a cost-and time-effective screening. cardiovascular side effects are critical in drug development and have frequently led to late-stage project terminations or even drug withdrawal from the market. physiologically relevant and predictive assays for cardiotoxicity are hence strongly demanded by the pharmaceutical industry. to identify a potential impact of test compounds on ventricular repolarization, typically a variety of ion channels in diverse heterologously expressing cells have to be investigated. Similar to primary cells, in vitro-generated stem cell-derived cardiomyocytes simultaneously express cardiac ion channels. thus, they more accurately represent the native situation compared with cell lines overexpressing only a single type of ion channel. the aim of this study was to determine if stem cell-derived cardiomyocytes are suited for use in an automated patch clamp system. the authors show recordings of cardiac ion currents as well as action potential recordings in readily available stem cell-derived cardiomyocytes. Besides monitoring inhibitory effects of reference compounds on typical cardiac ion currents, the authors revealed for the first time drug-induced modulation of cardiac action potentials in an automa...

Integrated platform for functional monitoring of biomimetic heart sheets derived from human pluripotent stem cells

“…cost and limited supply of hPSC-CMs, invasive characterization assays are undesirable. While there are setups for high-throughput measurements [22], these methods typically provide less content. Several non-invasive, bright field based, characterization techniques have recently been proposed.…”

Regulatory aspects of small molecule drugs for heart regeneration

“…Combining patient-specific iPSC-cardiomyocytes (iPSC-CM) with emerging highthroughput methods could create an extremely powerful system for the screening of potential novel therapies for these disease states. One such example is an automated patch clamp screening platform (e.g., SyncroPatch 96, Nanion Technologies, Germany), which has the ability to record data from 96 cells in parallel with a throughput of 5000 data points per day, allowing for the generation of full doseresponse curves from individual cells [95]. Recent advances in microelectrode arrays (MEAs) have also made it possible to adapt this technology to high-throughput-based assays.…”

Novel screening techniques for ion channel targeting drugs

“…Cell lines such as HEK and CHO cells are used with equal success rates and experiments can be performed at room or physiological temperature. 8 Additionally, primary cells such as human corneal epithelial cells, 9 stem cellderived cardiomyocytes 10 have been used successfully. As the data recorded is low noise and high resolution, single channel recordings in the cell attached mode on the cell membrane, and experiments recording ion channels reconstituted into lipid bilayers, are also routinely performed using the Port-a-Patch.…”

“…2,25 Included in this is the evaluation of the use of pluripotent stem cell applications for cardiovascular risk assessment. So far, the Patchliner has been used successfully to record from mouse embryonic stem cells (CorAt, Axiogenesis), 10 human iPS cells from CDI, 14,26 Axiogenesis (Cor.4U) 26 ), and Cellectis ( Fig. 2A, B).…”