Human pluripotent stem cell-derived cardiomyocytes align under cyclic strain when guided by cardiac fibroblasts

APL Bioengineering, vol. 6, pp. 046108

Abstract

The myocardium is a mechanically active tissue typified by anisotropy of the resident cells [cardiomyocytes (CMs) and cardiac fibroblasts (cFBs)] and the extracellular matrix (ECM). Upon ischemic injury, the anisotropic tissue is replaced by disorganized scar tissue, resulting in loss of coordinated contraction. Efforts to re-establish tissue anisotropy in the injured myocardium are hampered by a lack of understanding of how CM and/or cFB structural organization is affected by the two major physical cues inherent in the myocardium: ECM organization and cyclic mechanical strain. Herein, we investigate the singular and combined effect of ECM (dis)organization and cyclic strain in a two-dimensional human in vitro co-culture model of the myocardial microenvironment. We show that (an)isotropic ECM protein patterning can guide the orientation of CMs and cFBs, both in mono- and co-culture. Subsequent application of uniaxial cyclic strain—mimicking the local anisotropic deformation of beating myocardium—causes no effect when applied parallel to the anisotropic ECM. However, when cultured on isotropic substrates, cFBs, but not CMs, orient away from the direction of cyclic uniaxial strain (strain avoidance). In contrast, CMs show strain avoidance via active remodeling of their sarcomeres only when co-cultured with at least 30% cFBs. Paracrine signaling or N-cadherin-mediated communication between CMs and cFBs was no contributing factor. Our findings suggest that the mechanoresponsive cFBs provide structural guidance for CM orientation and elongation. Our study, therefore, highlights a synergistic mechanobiological interplay between CMs and cFBs in shaping tissue organization, which is of relevance for regenerating functionally organized myocardium.

Authors 7

1. Dylan Mostert

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology 1 , PO Box 513, 5600 MB Eindhoven, The Netherlands

   Institute for Complex Molecular Systems (ICMS) 2 , PO Box 513, 5600 MB Eindhoven, The Netherlands

2. Bart G. W. Groenen

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology 1 , PO Box 513, 5600 MB Eindhoven, The Netherlands

3. Leda Klouda

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology 1 , PO Box 513, 5600 MB Eindhoven, The Netherlands

4. Robert C. J. J. Passier Aachen Department of Anatomy and Embryology

   Leiden University · Leiden University Medical Center · University of Twente

   Affiliation as printed

   Department of Anatomy and Embryology, Leiden University Medical Centre 4 , Leiden, The Netherlands

   Department of Applied Stem Cell Technologies, University of Twente 3 , Enschede, The Netherlands

5. Marie‐José Goumans Aachen Department of Cell and Chemical Biology and Center for Biomedical Genetics

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Department of Cell and Chemical Biology and Center for Biomedical Genetics, Leiden University Medical Centre 5 , Leiden, The Netherlands

6. Nicholas A. Kurniawan

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology 1 , PO Box 513, 5600 MB Eindhoven, The Netherlands

   Institute for Complex Molecular Systems (ICMS) 2 , PO Box 513, 5600 MB Eindhoven, The Netherlands

7. Carlijn V. C. Bouten corresponding

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology 1 , PO Box 513, 5600 MB Eindhoven, The Netherlands

   Institute for Complex Molecular Systems (ICMS) 2 , PO Box 513, 5600 MB Eindhoven, The Netherlands