Conventional rigid 2D substrates cause complex contractile signals in monolayers of human induced pluripotent stem cell derived cardiomyocytes

Huethorst, E. , Mortensen, P., Simitev, R. D. , Gao, H. , Pohjolainen, L., Talman, V., Ruskoaho, H., Burton, F. L., Gadegaard, N. and Smith, G. L. (2022) Conventional rigid 2D substrates cause complex contractile signals in monolayers of human induced pluripotent stem cell derived cardiomyocytes. Journal of Physiology, 600(3), pp. 483-507. (doi: 10.1113/JP282228) (PMID:34761809)

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Abstract

Key points: Spatiotemporal contractility analysis of human induced pluripotent stem cell derived cardiomyocyte (hiPSC-CM) monolayers seeded on conventional, rigid surfaces (glass or plastic) revealed the presence of multiphasic contraction patterns across the monolayer with a high variability, despite action potentials recorded in the same areas being identical. These multiphasic patterns are not present in single cells, in detached monolayers or in monolayers seeded on soft substrates such as a hydrogel, where only ‘twitch’-like transients are observed. HiPSC-CM monolayers that display a high percentage of regions with multiphasic contraction have significantly increased contractile duration and a decreased lusotropic drug response. There is no indication that the multiphasic contraction patterns are associated with significant activation of the stress-activated NPPA or NPPB signalling pathways. A computational model of cell clusters supports the biological findings that the rigid surface and the differential cell-substrate adhesion underly multiphasic contractile behaviour of hiPSC-CMs. Abstract: Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) in monolayers interact mechanically via cell-cell and cell-substrate adhesion. Spatiotemporal features of contraction were analysed in hiPSC-CM monolayers 1) attached to glass or plastic (Young's modulus (E) >1 GPa), 2) detached (substrate-free) and 3) attached to a flexible collagen hydrogel (E = 22 kPa). The effects of isoprenaline on contraction were compared between rigid and flexible substrates. To clarify underlying mechanisms, further gene expression and computational studies were performed. HiPSC-CM monolayers exhibited multi-phasic contractile profiles on rigid surfaces in contrast to hydrogels, substrate-free cultures or single cells where only simple twitch-like time-courses were observed. Isoprenaline did not change the contraction profile on either surface, but its lusitropic and chronotropic effects were greater in hydrogel compared to glass. There was no significant difference between stiff and flexible substrates in regard to expression of the stress activated genes NPPA and NPPB. A computational model of cell clusters demonstrated similar complex contractile interactions on stiff substrates as a consequence of cell-to-cell functional heterogeneity. Rigid biomaterial surfaces give rise to unphysiological, multi-phasic contractions in hiPSC-CM monolayers. Flexible substrates are necessary for normal twitch-like contractility kinetics and interpretation of inotropic interventions.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Burton, Dr Francis and Simitev, Professor Radostin and Smith, Professor Godfrey and Gao, Dr Hao and Gadegaard, Professor Nikolaj and Mortensen, Mr Peter and Huethorst, Miss Eline
Authors: Huethorst, E., Mortensen, P., Simitev, R. D., Gao, H., Pohjolainen, L., Talman, V., Ruskoaho, H., Burton, F. L., Gadegaard, N., 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 > Biomedical Engineering
College of Science and Engineering > School of Engineering > Infrastructure and Environment
College of Science and Engineering > School of Mathematics and Statistics > Mathematics
Journal Name:Journal of Physiology
Publisher:Wiley
ISSN:0022-3751
ISSN (Online):1469-7793
Published Online:11 November 2021
Copyright Holders:Copyright © 2021 The Authors
First Published:First published in Journal of Physiology 600(3): 483-507
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
173470BHF 4-Year PhD Studentship (4th intake 2016 of 3rd Scheme)Rhian TouyzBritish Heart Foundation (BHF)FS/16/55/32731Institute of Cardiovascular & Medical Sciences
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