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

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

Abstract The human myocardium is a mechanically active tissue typified by anisotropic organization 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, eventually resulting in loss of coordinated contraction. Efforts to re-establish tissue anisotropy in the injured myocardium are hampered by a lack of understanding on 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 2D 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 were no contributing factors, but 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

   Eindhoven University of Technology, Department of Biomedical Engineering

   Institute for Complex Molecular Systems (ICMS)

   Eindhoven University of Technology, Department of Biomedical Engineering, PO Box 53, 5600 MB Eindhoven, The Netherlands

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

2. Bart G. W. Groenen

   Eindhoven University of Technology

   Affiliation as printed

   Eindhoven University of Technology, Department of Biomedical Engineering

   Eindhoven University of Technology, Department of Biomedical Engineering, PO Box 53, 5600 MB Eindhoven, The Netherlands

3. Leda Klouda

   Eindhoven University of Technology

   Affiliation as printed

   Eindhoven University of Technology, Department of Biomedical Engineering

   Eindhoven University of Technology, Department of Biomedical Engineering, PO Box 53, 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

   Leiden University Medical Centre, Department of Anatomy and Embryology

   University of Twente, Department of Applied Stem Cell Technologies

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

   University of Twente, Department of Applied Stem Cell Technologies, 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

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

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

6. Nicholas A. Kurniawan

   Eindhoven University of Technology

   Affiliation as printed

   Eindhoven University of Technology, Department of Biomedical Engineering

   Institute for Complex Molecular Systems (ICMS)

   Eindhoven University of Technology, Department of Biomedical Engineering, PO Box 53, 5600 MB Eindhoven, The Netherlands

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

7. Carlijn V. C. Bouten corresponding

   Eindhoven University of Technology

   Affiliation as printed

   Eindhoven University of Technology, Department of Biomedical Engineering

   Institute for Complex Molecular Systems (ICMS)

   Eindhoven University of Technology, Department of Biomedical Engineering, PO Box 53, 5600 MB Eindhoven, The Netherlands

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