Automated electrophysiological and pharmacological evaluation of human pluripotent stem cell-derived cardiomyocytes
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 harve...
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| Format: | Article |
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MAry Ann Liebert
2016
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| Online Access: | https://eprints.nottingham.ac.uk/37773/ |
| _version_ | 1848795530771038208 |
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| author | Rajamohan, Divya Kalra, Spandan Hoang, Minh Duc George, Vinoj Staniforth, Andrew Russell, Hugh Yang, Xuebin Denning, Chris |
| author_facet | Rajamohan, Divya Kalra, Spandan Hoang, Minh Duc George, Vinoj Staniforth, Andrew Russell, Hugh Yang, Xuebin Denning, Chris |
| author_sort | Rajamohan, Divya |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T19:33:33Z |
| format | Article |
| id | nottingham-37773 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:33:33Z |
| publishDate | 2016 |
| publisher | MAry Ann Liebert |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-377732020-05-04T17:41:57Z https://eprints.nottingham.ac.uk/37773/ 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 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. MAry Ann Liebert 2016-03-15 Article PeerReviewed Rajamohan, Divya, Kalra, Spandan, Hoang, Minh Duc, George, Vinoj, Staniforth, Andrew, Russell, Hugh, Yang, Xuebin and Denning, Chris (2016) Automated electrophysiological and pharmacological evaluation of human pluripotent stem cell-derived cardiomyocytes. Stem Cells and Development, 25 (6). pp. 439-452. ISSN 1557-8534 http://online.liebertpub.com/doi/10.1089/scd.2015.0253 doi:10.1089/scd.2015.0253 doi:10.1089/scd.2015.0253 |
| spellingShingle | Rajamohan, Divya Kalra, Spandan Hoang, Minh Duc George, Vinoj Staniforth, Andrew Russell, Hugh Yang, Xuebin Denning, Chris Automated electrophysiological and pharmacological evaluation of human pluripotent stem cell-derived cardiomyocytes |
| title | Automated electrophysiological and pharmacological evaluation of human pluripotent stem cell-derived cardiomyocytes |
| title_full | Automated electrophysiological and pharmacological evaluation of human pluripotent stem cell-derived cardiomyocytes |
| title_fullStr | Automated electrophysiological and pharmacological evaluation of human pluripotent stem cell-derived cardiomyocytes |
| title_full_unstemmed | Automated electrophysiological and pharmacological evaluation of human pluripotent stem cell-derived cardiomyocytes |
| title_short | Automated electrophysiological and pharmacological evaluation of human pluripotent stem cell-derived cardiomyocytes |
| title_sort | automated electrophysiological and pharmacological evaluation of human pluripotent stem cell-derived cardiomyocytes |
| url | https://eprints.nottingham.ac.uk/37773/ https://eprints.nottingham.ac.uk/37773/ https://eprints.nottingham.ac.uk/37773/ |