Electrophysiology of hiPSC-cardiomyocytes co-cultured with HEK cells expressing the inward rectifier channel

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)

[img] Text
244739.pdf - Published Version
Available under License Creative Commons Attribution.

6MB

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:Burton, Dr Francis and Da Silva Costa, Miss Ana and Simitev, Professor Radostin and Smith, Professor Godfrey and Gao, Dr Hao 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 > Institute of Cardiovascular and Medical Sciences
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

University Staff: Request a correction | Enlighten Editors: Update this record

Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
172141EPSRC Centre for Multiscale soft tissue mechanics with application to heart & cancerRaymond OgdenEngineering and Physical Sciences Research Council (EPSRC)EP/N014642/1M&S - Mathematics
303232EPSRC Centre for Multiscale soft tissue mechanics with MIT and POLIMI (SofTMech-MP)Xiaoyu LuoEngineering and Physical Sciences Research Council (EPSRC)EP/S030875/1M&S - Mathematics
308255The SofTMech Statistical Emulation and Translation HubDirk HusmeierEngineering and Physical Sciences Research Council (EPSRC)EP/T017899/1M&S - Statistics