In situ monolayer patch clamp of acutely stimulated human iPSC-derived cardiomyocytes promotes consistent electrophysiological responses to SK channel inhibition

• Published in: Scientific reports Vol. 14; no. 1; p. 3185
• Main Authors: Butler, Andrew S., Ascione, Raimondo, Marrion, Neil V., Harmer, Stephen C., Hancox, Jules C.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 07.02.2024; Nature Publishing Group; Nature Portfolio
• Subjects: 631/1647/1453; 631/532/2064/2158; 631/532/2440; Adult; Calcium channels; Calcium conductance; Cardiomyocytes; Cell Differentiation - genetics; Electrical stimuli; Heart; Heart Atria; Heterogeneity; Humanities and Social Sciences; Humans; Induced Pluripotent Stem Cells - metabolism; Ion channels; multidisciplinary; Myocytes, Cardiac - metabolism; Phenotypes; Pluripotency; Potassium channels; Potassium conductance; Retinoic acid; Science; Science (multidisciplinary); Stem cells
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-53571-6

Human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) represent an in vitro model of cardiac function. Isolated iPSC-CMs, however, exhibit electrophysiological heterogeneity which hinders their utility in the study of certain cardiac currents. In the healthy adult heart, the current mediated by small conductance, calcium-activated potassium (SK) channels (I SK ) is atrial-selective. Functional expression of I SK within atrial-like iPSC-CMs has not been explored thoroughly. The present study therefore aimed to investigate atrial-like iPSC-CMs as a model system for the study of I SK . iPSCs were differentiated using retinoic acid (RA) to produce iPSC-CMs which exhibited an atrial-like phenotype (RA-iPSC-CMs). Only 18% of isolated RA-iPSC-CMs responded to SK channel inhibition by UCL1684 and isolated iPSC-CMs exhibited substantial cell-to-cell electrophysiological heterogeneity. This variability was significantly reduced by patch clamp of RA-iPSC-CMs in situ as a monolayer (iPSC-ML). A novel method of electrical stimulation was developed to facilitate recording from iPSC-MLs via In situ M onolayer P atch clamp of A cutely S timulated iPSC- C Ms (IMPASC). Using IMPASC, > 95% of iPSC-MLs could be paced at a 1 Hz. In contrast to isolated RA-iPSC-CMs, 100% of RA-iPSC-MLs responded to UCL1684, with APD 50 being prolonged by 16.0 ± 2.0 ms (p < 0.0001; n  = 12). These data demonstrate that in conjunction with IMPASC, RA-iPSC-MLs represent an improved model for the study of I SK . IMPASC may be of wider value in the study of other ion channels that are inconsistently expressed in isolated iPSC-CMs and in pharmacological studies.