Recording of multiple ion current components and action potentials in human induced pluripotent stem cell-derived cardiomyocytes via automated patch-clamp

Mann, Stefan A.; Heide, Juliane; Knott, Thomas; Airini, Razvan; Epureanu, Florin Bogdan; Deftu, Alexandru‐Florian; Deftu, Antonia-Teona; Radu, Beatrice Mihaela; Amuzescu, Bogdan

doi:10.1016/j.vascn.2019.106599

Cited by 22 publications

(31 citation statements)

References 89 publications

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“…In fact, the mean capacitance of iCell cardiomyocytes, ventricular Pluricytes and atrial Pluricytes was comparable (≈ 20-25 pF) and smaller than the mean capacitance of primary human ventricular cardiomyocytes (≈ 123 pF). This finding is in agreement with earlier reports (see collected data of human cardiomyocytes in (Polak & Fijorek, 2012) and data for hiPSC-CMs (Mann et al, 2019). Ventricular Pluricytes showed a slightly higher capacitance than atrial Pluricytes.…”

Section: Discussionsupporting

confidence: 94%

Ion channel expression and electrophysiology of singular human (primary and induced pluripotent stem cell derived) cardiomyocytes

Schmid

Abi‐Gerges

Zellner

et al. 2021

Preprint

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Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and primary human cardiomyocytes are used for in vitro cardiac safety testing. hiPSC-CMs have been associated with a vast heterogeneity regarding single-cell morphology, beating behavior and action potential duration, prompting a systematic analysis of single-cell characteristics. Previously published hiPSC-CM studies revealed action potentials with nodal-, atrial- or ventricular-like morphology, although ion channel expression of singular hiPSC-CMs is not fully understood. Other studies used single-cell RNA-sequencing, however, these studies did not extensively focus on expression patterns of cardiac ion channels or failed to detect ion channel transcripts. Thus, the current study used a single-cell patch-clamp-RT-qPCR approach to get insights into single-cell electrophysiology (capacitance, action potential duration at 90% of repolarization, upstroke velocity, spontaneous beat rate, and sodium-driven fast inward current) and ion channel expression (HCN4, CACNA1G, CACNA1D, KCNA5, KCNJ4, SCN5A, KCNJ2, CACNA1D, and KCNH2), the combination of both within individual cells, and their correlations in single cardiomyocytes. We used commercially available hiPSC-CMs (iCell cardiomyocytes, atrial and ventricular Pluricytes) and primary human adult atrial and ventricular cardiomyocytes. Recordings of electrophysiological parameters revealed differences between the cell groups and variation within the hiPSC-CMs groups as well as within primary ventricular cardiomyocytes. Expression analysis on mRNA level showed no-clear-cut discrimination between primary cardiac subtypes and revealed both similarities and differences between all cell groups. Higher expression of atrial-associated ion channels in primary atrial cardiomyocytes and atrial Pluricytes compared to their ventricular counterpart indicates a successful chamber-specific hiPSC differentiation. Interpretation of correlations between the single-cell parameters was challenging, as the total data set is complex, particularly for parameters depending on multiple processes, like the spontaneous beat rate. Yet, for example, expression of SCN5A correlated well with the fast inward current amplitude for all three hiPSC-CM groups. To further enhance our understanding of the physiology and composition of the investigated hiPSC-CMs, we compared beating and non-beating cells and assessed distributions of single-cell data. Investigating the single-cell phenotypes of hiPSC-CMs revealed a combination of attributes which may be interpreted as a mixture of traits of different adult cardiac cell types: (i) nodal-related pacemaking attributes are spontaneous generation of action potentials and high HCN4 expression; and (ii) non-nodal attributes: cells have a prominent INa-driven fast inward current, a fast upstroke velocity and a high expression of SCN5A. In conclusion, the combination of nodal- and non-nodal attributes in single hiPSC-CMs may hamper the interpretation of drug effects on complex electrophysiological parameters like beat rate and action potential duration. However, the proven expression of specific ion channels enables the evaluation of drug effects on ionic currents in a more realistic environment than in recombinant systems.

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Section: Discussionsupporting

confidence: 94%

Ion channel expression and electrophysiology of singular human (primary and induced pluripotent stem cell derived) cardiomyocytes

Schmid

Abi‐Gerges

Zellner

et al. 2021

Preprint

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“…Human iPSC derived cardiomyocytes are genetically isomorphous with human ventricular cells, have action potentials that resemble those of developing foetal human ventricular myocytes, and have the same, but quantitatively different expression profile of membrane ionic channels ( Mann et al, 2019 ; Kernik at al., 2019 ). Figure 6 illustrates the action potential responses to pacing with a BCL of 10 s of an isolated hiPSC cardiomyocyte at room temperature to CORM-2.…”

Section: Resultsmentioning

confidence: 99%

“…For the population of cell models, with 5% variability in all the membrane maximal conductance parameters, the effect of CO on the endocardial models is to prolong the APD, and increase its variability, with EADS produced in <1% of the variant models. In the epicardial cell models the APD and its variability are increased producing EADs and a bimodal distribution of Human iPSC derived cardiomyocytes are genetically isomorphous with human ventricular cells, have action potentials that resemble those of developing foetal human ventricular myocytes, and have the same, but quantitatively different expression profile of membrane ionic channels (Mann et al, 2019;Kernik at al., 2019). Figure 6 illustrates the action potential responses to pacing with a BCL of 10 s of an isolated hiPSC cardiomyocyte at room temperature to CORM-2.…”

Section: Carbon Monoxide Prolongs Action Potential Duration and Induced Early After-depolarizations In Guinea Pig Ventricular Myocytesmentioning

confidence: 99%

Deterministic and Stochastic Cellular Mechanisms Contributing to Carbon Monoxide Induced Ventricular Arrhythmias

et al. 2021

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Chronic exposure to low levels of Carbon Monoxide is associated with an increased risk of cardiac arrhythmia. Microelectrode recordings from rat and guinea pig single isolated ventricular myocytes exposed to CO releasing molecule CORM-2 and excited at 0.2/s show repolarisation changes that develop over hundreds of seconds: action potential prolongation by delayed repolarisation, EADs, multiple EADs and oscillations around the plateau, leading to irreversible repolarisation failure. The measured direct effects of CO on currents in these cells, and ion channels expressed in mammalian systems showed an increase in prolonged late Na+, and a decrease in the maximal T- and L-type Ca++. peak and late Na+, ultra-rapid delayed, delayed rectifier, and the inward rectifier K+ currents. Incorporation of these CO induced changes in maximal currents in ventricular cell models; (Gattoni et al., J. Physiol., 2016, 594, 4193–4224) (rat) and (Luo and Rudy, Circ. Res., 1994, 74, 1071–1096) (guinea-pig) and human endo-, mid-myo- and epi-cardial (O’Hara et al., PLoS Comput. Biol., 2011, 7, e1002061) models, by changes in maximal ionic conductance reproduces these repolarisation abnormalities. Simulations of cell populations with Gaussian distributions of maximal conductance parameters predict a CO induced increase in APD and its variability. Incorporation of these predicted CO induced conductance changes in human ventricular cell electrophysiology into ventricular tissue and wall models give changes in indices for the probability of the initiation of re-entrant arrhythmia.

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“…Microfluidic devices are considered in translational medicine studies by institutions such as the U.S. Food and Drug Administration (FDA) [ 176 ], which supports the initiative to develop such chips for clinical trials to help personalized therapies. A prerequisite of any new drug development is to overcome several important drug safety tests, which mandatory include its interaction with BBB (overcoming or not the barrier) [ 177 ] and its proarrhythmogenic risk [ 178 , 179 ]. Other safety tests, such as renal or liver toxicity, may also be considered.…”

Section: Loc Microdevices In Translational Medicine With Impact In Neurological Disordersmentioning

confidence: 99%

Lab-on-a-Chip Platforms as Tools for Drug Screening in Neuropathologies Associated with Blood–Brain Barrier Alterations

et al. 2021

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Lab-on-a-chip (LOC) and organ-on-a-chip (OOC) devices are highly versatile platforms that enable miniaturization and advanced controlled laboratory functions (i.e., microfluidics, advanced optical or electrical recordings, high-throughput screening). The manufacturing advancements of LOCs/OOCs for biomedical applications and their current limitations are briefly discussed. Multiple studies have exploited the advantages of mimicking organs or tissues on a chip. Among these, we focused our attention on the brain-on-a-chip, blood–brain barrier (BBB)-on-a-chip, and neurovascular unit (NVU)-on-a-chip applications. Mainly, we review the latest developments of brain-on-a-chip, BBB-on-a-chip, and NVU-on-a-chip devices and their use as testing platforms for high-throughput pharmacological screening. In particular, we analyze the most important contributions of these studies in the field of neurodegenerative diseases and their relevance in translational personalized medicine.

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Recording of multiple ion current components and action potentials in human induced pluripotent stem cell-derived cardiomyocytes via automated patch-clamp

Cited by 22 publications

References 89 publications

Ion channel expression and electrophysiology of singular human (primary and induced pluripotent stem cell derived) cardiomyocytes

Ion channel expression and electrophysiology of singular human (primary and induced pluripotent stem cell derived) cardiomyocytes

Deterministic and Stochastic Cellular Mechanisms Contributing to Carbon Monoxide Induced Ventricular Arrhythmias

Lab-on-a-Chip Platforms as Tools for Drug Screening in Neuropathologies Associated with Blood–Brain Barrier Alterations

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