0aef4e9ddb5a54532efdff6c9ec1a520 334587.pdf 34748d1f2b5ac447e28fcdd6f6014320672eee62 334587.pdf d0d1d52a78519d579f54ff7733988ab2be60941426f5f6af04bb1d94e4524ba8 334587.pdf Title: Electrophysiological Phenotyping of hiPSC-Derived Atrial Cardiomyocytes Using Automated Patch-Clamp: A Platform for Studying Atrial Inherited Arrhythmias Subject: Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) represent a robust platform for modelling inherited cardiac disorders. Comparative analysis of ion channel activity in patient-specific and isogenic control lines provides critical insights into the molecular mechanisms underlying channelopathies and arrhythmias. Atrial-specific hiPSC-CMs (hiPSC-aCMs) exhibit distinct electrophysiological properties governed by unique ion channel expression profiles, underscoring the need for optimized methodologies to record atrial ionic currents accurately. Here, we characterized the electrophysiological features of hiPSC-aCMs using the Nanion Patchliner automated patch-clamp system. An optimized cell dissociation protocol was developed to enhance cell integrity and seal formation, while tailored intra- and extracellular solutions were employed to isolate specific ionic currents. Using this approach, we reliably recorded major atrial currents, including the sodium current (INa), L-type calcium current (ICaL), transient outward potassium current (Ito), ultrarapid component of the delayed rectifier current (IKur), small-conductance calcium-activated potassium current (ISK), and pacemaker funny current (If). The resulting current profiles were reproducible and consistent with those observed in native atrial cardiomyocytes. These findings establish the feasibility of the automated electrophysiological characterization of ion channels in hiPSC-aCMs. This platform enables more efficient investigation of pathogenic variants and facilitates the development of targeted therapeutics for atrial arrhythmias and related channelopathies. Keywords: automated patch-clamp; cardiac ion channels; human-induced pluripotent stem cell-derived atrial cardiomyocytes (hiPSC-aCMs); arrhythmia Author: Verónica Jiménez-Sábado, Hosna Babini, Peter C. Ruben, Eric A. Accili, Thomas W. Claydon, Leif Hove-Madsen and Glen F. Tibbits Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Sat Dec 6 10:38:36 2025 CET ModDate: Sat Dec 6 10:43:29 2025 CET Custom Metadata: yes Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 21 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2592988 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- LUVXVU+CMSY10 Type 1 Builtin yes yes yes 141 0 DRHGYZ+URWPalladioL-Bold Type 1 Custom yes yes yes 142 0 BAKGOS+URWPalladioL-Roma Type 1 Custom yes yes yes 143 0 YCFZZZ+URWPalladioL-Ital Type 1 Custom yes yes yes 144 0 OKGWPZ+EURM10 Type 1 Builtin yes yes yes 250 0 ECFSDQ+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 230 0 SKGYRO+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 231 0 ZFQOLA+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 232 0 NSQUZY+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 243 0 ECFSDQ+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 244 0 SKGYRO+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 245 0 ZFQOLA+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 246 0 SPXLNH+PazoMath Type 1 Builtin yes yes yes 299 0 NSQUZY+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 321 0 SKGYRO+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 322 0 ZFQOLA+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 323 0 ECFSDQ+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 324 0 NSQUZY+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 344 0 SKGYRO+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 345 0 ZFQOLA+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 346 0 NSQUZY+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 377 0 FOSYRO+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 378 0 SKGYRO+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 379 0 ZFQOLA+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 380 0 NSQUZY+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 403 0 FOSYRO+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 404 0 SKGYRO+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 405 0 ZFQOLA+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 406 0 NSQUZY+PalatinoLinotype-Bold TrueType WinAnsi yes yes yes 462 0 ECFSDQ+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 463 0 SKGYRO+PalatinoLinotype-Roman TrueType WinAnsi yes yes yes 464 0 ZFQOLA+PalatinoLinotype-Italic TrueType WinAnsi yes yes yes 465 0 BAZSKP+VnURWPalladioL Type 1 Custom yes yes yes 678 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2026-08-01 05:19:32 CEST RepresentationInformation: 334587.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2026-07-31 10:21:34 CEST Size: 2592988 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 871 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Materials and Methods Children: Item: Title: Differentiation of hiPSC into Atrial Cardiomyocytes Item: Title: Preparation of hiPSC-aCMs for Automated Patch-Clamp Experiments Item: Title: Automated Patch-Clamp Technique Children: Item: Title: Solutions and Specific Protocols Used for Electrophysiological Recordings Item: Title: Measurement of Na+ Current Item: Title: Measurement of L-Type Calcium Current Item: Title: Measurement of Major Atrial Potassium Repolarizing Currents Item: Title: Measurement of Small-Conductance Calcium-Activated Potassium Current Item: Title: Measurement of Funny Current Item: Title: Data Analysis Children: Item: Title: Statistical Analysis Item: Title: Mathematical Analysis Item: Title: Results Children: Item: Title: Sodium Current Characterization in hiPSC-aCMs Item: Title: L-Type Calcium Current Characterization in hiPSC-aCMs Item: Title: Characterization of the Major Atrial K+ Repolarizing Currents in hiPSC-aCMs Item: Title: Apamin-Sensitive SK Current Characterization in hiPSC-aCMs Item: Title: Funny Current Characterization in hiPSC-aCMs Item: Title: Discussion Item: Title: Conclusions and Clinical Relevance Item: Title: References Info: Title: Electrophysiological Phenotyping of hiPSC-Derived Atrial Cardiomyocytes Using Automated Patch-Clamp: A Platform for Studying Atrial Inherited Arrhythmias Author: Verónica Jiménez-Sábado, Hosna Babini, Peter C. Ruben, Eric A. Accili, Thomas W. Claydon, Leif Hove-Madsen and Glen F. Tibbits Subject: Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) represent a robust platform for modelling inherited cardiac disorders. Comparative analysis of ion channel activity in patient-specific and isogenic control lines provides critical insights into the molecular mechanisms underlying channelopathies and arrhythmias. Atrial-specific hiPSC-CMs (hiPSC-aCMs) exhibit distinct electrophysiological properties governed by unique ion channel expression profiles, underscoring the need for optimized methodologies to record atrial ionic currents accurately. Here, we characterized the electrophysiological features of hiPSC-aCMs using the Nanion Patchliner automated patch-clamp system. An optimized cell dissociation protocol was developed to enhance cell integrity and seal formation, while tailored intra- and extracellular solutions were employed to isolate specific ionic currents. Using this approach, we reliably recorded major atrial currents, including the sodium current (INa), L-type calcium current (ICaL), transient outward potassium current (Ito), ultrarapid component of the delayed rectifier current (IKur), small-conductance calcium-activated potassium current (ISK), and pacemaker funny current (If). The resulting current profiles were reproducible and consistent with those observed in native atrial cardiomyocytes. These findings establish the feasibility of the automated electrophysiological characterization of ion channels in hiPSC-aCMs. This platform enables more efficient investigation of pathogenic variants and facilitates the development of targeted therapeutics for atrial arrhythmias and related channelopathies. 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