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Automated electrophysiological and pharmacological evaluation of human pluripotent stem cell-derived cardiomyocytes

Rajamohan, Divya; Kalra, Spandan; Hoang, Minh Duc; George, Vinoj; Staniforth, Andrew; Russell, Hugh; Yang, Xuebin; Denning, Chris

Authors

Divya Rajamohan

Spandan Kalra

Minh Duc Hoang

Vinoj George

Andrew Staniforth

Hugh Russell

Xuebin Yang

CHRIS DENNING chris.denning@nottingham.ac.uk
Professor of Stem Cell Biology



Abstract

Automated planar patch clamp systems are widely used in drug evaluation studies because of their ability to provide accurate, reliable, and reproducible data in a high-throughput manner. Typically, CHO and HEK tumorigenic cell lines overexpressing single ion channels are used since they can be harvested as high-density, homogenous, single-cell suspensions. While human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are physiologically more relevant, these cells are fragile, have complex culture requirements, are inherently heterogeneous, and are expensive to produce, which has restricted their use on automated patch clamp (APC) devices. Here, we used high efficiency differentiation protocols to produce cardiomyocytes from six different hPSC lines for analysis on the Patchliner (Nanion Technologies GmbH) APC platform. We developed a two-step cell preparation protocol that yielded cell catch rates and whole-cell breakthroughs of ∼80%, with ∼40% of these cells allowing electrical activity to be recorded. The protocol permitted formation of long-lasting (>15 min), high quality seals (>2 GΩ) in both voltage- and current-clamp modes. This enabled density of sodium, calcium, and potassium currents to be evaluated, along with dose–response curves to their respective channel inhibitors, tetrodotoxin, nifedipine, and E-4031. Thus, we show the feasibility of using the Patchliner platform for automated evaluation of the electrophysiology and pharmacology of hPSC-CMs, which will enable considerable increase in throughput for reliable and efficient drug evaluation.

Citation

Rajamohan, D., Kalra, S., Hoang, M. D., George, V., Staniforth, A., Russell, H., …Denning, C. (2016). Automated electrophysiological and pharmacological evaluation of human pluripotent stem cell-derived cardiomyocytes. Stem Cells and Development, 25(6), https://doi.org/10.1089/scd.2015.0253

Journal Article Type Article
Acceptance Date Jan 18, 2016
Online Publication Date Feb 23, 2016
Publication Date Mar 15, 2016
Deposit Date Oct 25, 2016
Publicly Available Date Oct 25, 2016
Journal Stem Cells and Development
Print ISSN 1547-3287
Electronic ISSN 1557-8534
Publisher Mary Ann Liebert
Peer Reviewed Peer Reviewed
Volume 25
Issue 6
DOI https://doi.org/10.1089/scd.2015.0253
Public URL http://eprints.nottingham.ac.uk/id/eprint/37773
Publisher URL http://online.liebertpub.com/doi/10.1089/scd.2015.0253
Copyright Statement Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0

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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0





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