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A modular method for high-throughput measurement of ion channel currents in cardiac myocytes

Nature Protocols (2026) Cite this article

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Abstract

The patch-clamp technique offers unparalleled insight into the electrical and biophysical behavior of excitable cells. However, it is a slow and low-throughput method that typically requires cells to be measured one by one. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) are regularly subjected to this technique to unravel the molecular mechanisms of cardiac diseases. Their use in direct patient treatment and successful drug development has been limited due to the lack of applicable high-throughput patch-clamp methods suited to successful hiPSC-CM measurement. Here we present a protocol employing a patch-clamp robot that addresses these limitations by using planar patch-clamp technology. We outline how to collect and handle hiPSC-CM for these experiments, along with optimized patch-clamp protocols for direct functional measurement of major cardiac ion channels including Kir2.1, NaV1.5, CaV1.2, Kv11.1 and Kir3.1/3.4. We further explain how the liquid-handling properties of this setup allow multiple patch-clamp protocols to be combined in sequence while the cell remains in whole-cell configuration. This allows for over a hundred-fold increase in functional data acquisition. These procedures can be carried out within 1 d by both skilled and non-electrophysiologists; however, some experience in cell culture and handling is required. Overall, this protocol enhances fast and reliable functional characterization of hiPSCl-CM and may increase their applicability for rapid and safe drug development.

Key points

  • Human induced pluripotent stem cell-derived cardiomyocytes are useful constructs in the study of cardiac disease mechanisms. This protocol highlights how to extract broad electrophysiological features from these constructs in high throughput.

  • Typical patch-clamp measurements suffer from low throughput and represent a major bottleneck in disease modeling and drug development. This protocol removes this limitation through the combination of simultaneous population patch-clamp and robotic liquid handling.

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Fig. 1: Method overview for high-throughput characterization of hiPSC-CM electrophysiological parameters.
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Fig. 2: External solution liquid handling.
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Fig. 3: Overview of hiPSC-CM used in this work.
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Fig. 4: Extraction of desired readouts during general and organizational analysis steps using an INa experiment example.
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Fig. 5: Broad functional ion channel characterization in hiPSC-CM using high-throughput patch-clamp.
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Fig. 6: Technical overview of high-throughput electrophysiology combined with hiPSC-CM.
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Fig. 7: Flexible compound application and screening using high-throughput electrophysiology.
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Data availability

The main data discussed in this protocol are available from the supporting primary research papers.

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Acknowledgements

We thank S. Kestel for excellent technical help and M. Dilaj for excellent secretarial assistance. This project was supported by grants from the Deutsche Forschungsgemeinschaft (DFG, VO 1568/3-1, VO1568/4-1, VO1568/3-2, VO1568/6-1, SFB1002 project A13, and from the German Center for Cardiovascular Research (DZHK, 81×4300102, ‘DNAfix’) to N.V. This project was further supported by a grant from the DFG under Germany’s Excellence Strategy (EXC 2067/1—390729940) to N.V. and T.M. Last, this project was supported by the Else Kröner Fresenius Foundation through the Else Kröner Fresenius Center for Optogenetic Therapies for T.M.

Author information

Author notes

  1. These authors contributed equally: Fitzwilliam Seibertz, Izzatulo Sobitov.

Authors and Affiliations

  1. Institute of Pharmacology and Toxicology, University Medical Center Göttingen, Georg-August University Göttingen, Göttingen, Germany

    Fitzwilliam Seibertz, Izzatulo Sobitov, Marcus L. Gerloff, Aiste Liutkute, Funsho E. Fakuade & Niels Voigt

  2. DZHK (German Center for Cardiovascular Research), partner site Lower Saxony, Göttingen, Germany

    Fitzwilliam Seibertz, Izzatulo Sobitov, Marcus L. Gerloff, Aiste Liutkute, Constanze Schmidt, Funsho E. Fakuade & Niels Voigt

  3. Cluster of Excellence ‘Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells’ (MBExC), University of Göttingen, Göttingen, Germany

    Fitzwilliam Seibertz, Izzatulo Sobitov, Aiste Liutkute, Alexey Alekseev, Thomas Mager, Funsho E. Fakuade & Niels Voigt

  4. Department of Cardiology and Pneumology, University Medicine Göttingen, Göttingen, Germany

    Izzatulo Sobitov & Constanze Schmidt

  5. Institute for Auditory Neuroscience and InnerEarLab, University Medical Center Göttingen, Göttingen, Germany

    Alexey Alekseev & Thomas Mager

  6. Advanced Optogenes Group, Institute for Auditory Neuroscience, University Medical Center Göttingen, Göttingen, Germany

    Alexey Alekseev & Thomas Mager

  7. Else Kröner Fresenius Center for Optogenetic Therapies, University Medical Center Göttingen, Göttingen, Germany

    Alexey Alekseev & Thomas Mager

  8. Department of Cardiothoracic and Vascular Surgery, University Medical Center Göttingen, Göttingen, Germany

    Funsho E. Fakuade

Authors
  1. Fitzwilliam Seibertz
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  8. Funsho E. Fakuade
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  9. Niels Voigt
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Contributions

F.S., I.S., A.A., T.M., F.F., C.S. and N.V. designed and planned experiments. F.S., I.S., M.G. and A.A. analyzed the data. F.S., I.S., M.G., A.L. and A.A. established the methods and performed the experiments. F.S. and N.V. wrote the manuscript. All authors read and approved the manuscript.

Corresponding author

Correspondence to Niels Voigt.

Ethics declarations

Competing interests

F.S. became employed by Nanion Technologies GmbH (Germany) near the conclusion of this project. The other authors declare no competing interests.

Peer review

Peer review information

Nature Protocols thanks Henry Colecraft, who co-reviewed with Sri Karthika Shangmugam; and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Key references

Seibertz, F. et al. Commun. Biol. 5, 969 (2022): https://doi.org/10.1038/s42003-022-03871-2

Kyriakopoulou, E. et al. Nat. Cardiovasc. Res. 2, 1262–1276 (2023): https://doi.org/10.1038/s44161-023-00378-9

Seibertz, F. et al. Cardiovasc. Res. 119, 2623–2637 (2023): https://doi.org/10.1093/cvr/cvad143

Seibertz, F. et al. Basic Res. Cardiol. 118, 14 (2023): https://doi.org/10.1007/s00395-022-00973-0

Fakuade, F. E. et al. Circulation 150, 544–559 (2024): https://doi.org/10.1161/CIRCULATIONAHA.123.066577

Supplementary information

Supplementary Information (download PDF )

Supplementary Methods 1–5, Figs. 1 and 2 and Table 1.

Reporting Summary (download PDF )

Supplementary Software 1 (download XLSX )

Example organizational analysis file for INa data.

Supplementary Software 2

File containing code for automatic extraction of desired data from result tables.

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Seibertz, F., Sobitov, I., Gerloff, M.L. et al. A modular method for high-throughput measurement of ion channel currents in cardiac myocytes. Nat Protoc (2026). https://doi.org/10.1038/s41596-026-01351-z

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