Da Silva Costa, A. , Mortensen, P., Hortigon-Vinagre, M. P., van der Heyden, M. A. G., Burton, F. L., Gao, H. , Simitev, R. D. and Smith, G. L. (2021) Electrophysiology of hiPSC-cardiomyocytes co-cultured with HEK cells expressing the inward rectifier channel. International Journal of Molecular Sciences, 22(12), 6621. (doi: 10.3390/ijms22126621)
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Abstract
The immature electrophysiology of human-induced pluripotent stem cell-derived cardiomyocytes (hiCMs) complicates their use for therapeutic and pharmacological purposes. An insufficient inward rectifying current (IK1) and the presence of a funny current (if) cause spontaneous electrical activity. This study tests the hypothesis that the co-culturing of hiCMs with a human embryonic kidney (HEK) cell-line expressing the Kir2.1 channel (HEK-IK1) can generate an electrical syncytium with an adult-like cardiac electrophysiology. The mechanical activity of co-cultures using different HEK-IK1:hiCM ratios was compared with co-cultures using wildtype (HEK–WT:hiCM) or hiCM alone on days 3–8 after plating. Only ratios of 1:3 and 1:1 showed a significant reduction in spontaneous rate at days 4 and 6, suggesting that IK1 was influencing the electrophysiology. Detailed analysis at day 4 revealed an increased incidence of quiescent wells or sub-areas. Electrical activity showed a decreased action potential duration (APD) at 20% and 50%, but not at 90%, alongside a reduced amplitude of the aggregate AP signal. A computational model of the 1:1 co-culture replicates the electrophysiological effects of HEK–WT. The addition of the IK1 conductance reduced the spontaneous rate and APD20, 50 and 90, and minor variation in the intercellular conductance caused quiescence. In conclusion, a 1:1 co-culture HEK-IK1:hiCM caused changes in electrophysiology and spontaneous activity consistent with the integration of IK1 into the electrical syncytium. However, the additional electrical effects of the HEK cell at 1:1 increased the possibility of electrical quiescence before sufficient IK1 was integrated into the syncytium.
| Item Type: | Articles |
|---|---|
| Additional Information: | Funding: The research of R.D.S., H.G. and G.S. was funded by the UK Engineering and Physical Sciences Research Council, grant numbers EP/N014642/1, EP/S030875/1 and EP/T017899/1. A.D.S.C. was in receipt of an EPSRC training award number: 2053958. |
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Simitev, Professor Radostin and Smith, Professor Godfrey and Da Silva Costa, Dr Ana and Gao, Dr Hao and Burton, Dr Francis and Mortensen, Mr Peter |
| Creator Roles: | Da Silva Costa, A.Methodology, Formal analysis, Investigation, Data curation, Writing – original draft, Writing – review and editing, Visualization Mortensen, P.Methodology, Software, Formal analysis, Investigation, Data curation, Writing – original draft Burton, F. L.Software, Writing – review and editing Gao, H.Software, Resources, Writing – review and editing, Supervision, Funding acquisition Simitev, R. D.Software, Validation, Investigation, Writing – review and editing, Supervision, Funding acquisition Smith, G. L.Conceptualization, Validation, Investigation, Resources, Writing – original draft, Writing – review and editing, Supervision, Funding acquisition |
| Authors: | Da Silva Costa, A., Mortensen, P., Hortigon-Vinagre, M. P., van der Heyden, M. A. G., Burton, F. L., Gao, H., Simitev, R. D., and Smith, G. L. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Cardiovascular & Metabolic Health College of Science and Engineering > School of Engineering > Infrastructure and Environment College of Science and Engineering > School of Mathematics and Statistics > Mathematics |
| Journal Name: | International Journal of Molecular Sciences |
| Publisher: | MDPI |
| ISSN: | 1661-6596 |
| ISSN (Online): | 1422-0067 |
| Published Online: | 21 June 2021 |
| Copyright Holders: | Copyright © 2021 by the authors |
| First Published: | First published in International Journal of Molecular Sciences 22(12):e6621 |
| Publisher Policy: | Reproduced under a Creative Commons Licence |
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