Human-iPSC-Derived Cardiac Stromal Cells Enhance Maturation in 3D Cardiac Microtissues and Reveal Non-cardiomyocyte Contributions to Heart Disease.
Elisa Giacomelli, Viviana Meraviglia, Giulia Campostrini, Amy Cochrane, Xu Cao, Ruben W J van Helden, Ana Krotenberg Garcia, Maria Mircea, Sarantos Kostidis, Richard P Davis, Berend J van Meer, Carolina R Jost, Abraham J Koster, Hailiang Mei, David G Míguez, Aat A Mulder, Mario Ledesma-Terrón, Giulio Pompilio, Luca Sala, Daniela C F Salvatori, Roderick C Slieker, Elena Sommariva, Antoine A F de Vries, Martin Giera, Stefan Semrau, Leon G J Tertoolen, Valeria V Orlova, Milena Bellin, Christine L Mummery
Author Information
Elisa Giacomelli: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Viviana Meraviglia: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Giulia Campostrini: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Amy Cochrane: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Xu Cao: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Ruben W J van Helden: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Ana Krotenberg Garcia: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Maria Mircea: Leiden Institute of Physics, Leiden University, 2333 Leiden, the Netherlands.
Sarantos Kostidis: Center for Proteomics and Metabolomics, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Richard P Davis: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Berend J van Meer: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Carolina R Jost: Department of Cell and Chemical Biology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Abraham J Koster: Department of Cell and Chemical Biology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Hailiang Mei: Sequencing Analysis Support Core, Leiden University Medical Center, 2333 Leiden, the Netherlands.
David G Míguez: Centro de Biologia Molecular Severo Ochoa, Departamento de Física de la Materia Condensada, Instituto Nicolas Cabrera and Condensed Matter Physics Center (IFIMAC), Universidad Autónoma de Madrid, 28049 Madrid, Spain.
Aat A Mulder: Department of Cell and Chemical Biology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Mario Ledesma-Terrón: Centro de Biologia Molecular Severo Ochoa, Departamento de Física de la Materia Condensada, Instituto Nicolas Cabrera and Condensed Matter Physics Center (IFIMAC), Universidad Autónoma de Madrid, 28049 Madrid, Spain.
Giulio Pompilio: Vascular Biology and Regenerative Medicine Unit, Centro Cardiologico Monzino IRCCS, 20138 Milan, Italy; Department of Clinical Sciences and Community Health, Università degli Studi di Milano, 20122 Milan, Italy.
Luca Sala: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Daniela C F Salvatori: Central Laboratory Animal Facility, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Roderick C Slieker: Department of Cell and Chemical Biology, Leiden University Medical Center, 2333 Leiden, the Netherlands; Department of Epidemiology and Biostatistics, Amsterdam Public Health Institute, VU University Medical Center, 1007 Amsterdam, the Netherlands.
Elena Sommariva: Vascular Biology and Regenerative Medicine Unit, Centro Cardiologico Monzino IRCCS, 20138 Milan, Italy.
Antoine A F de Vries: Department of Cardiology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Martin Giera: Center for Proteomics and Metabolomics, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Stefan Semrau: Leiden Institute of Physics, Leiden University, 2333 Leiden, the Netherlands.
Leon G J Tertoolen: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands.
Valeria V Orlova: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands. Electronic address: v.orlova@lumc.nl.
Milena Bellin: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands; Department of Biology, University of Padua, 35121 Padua, Italy; Veneto Institute of Molecular Medicine, 35129 Padua, Italy. Electronic address: m.bellin@lumc.nl.
Christine L Mummery: Department of Anatomy and Embryology, Leiden University Medical Center, 2333 Leiden, the Netherlands; Department of Applied Stem Cell Technologies, University of Twente, 7500 Enschede, the Netherlands. Electronic address: c.l.mummery@lumc.nl.
Cardiomyocytes (CMs) from human induced pluripotent stem cells (hiPSCs) are functionally immature, but this is improved by incorporation into engineered tissues or forced contraction. Here, we showed that tri-cellular combinations of hiPSC-derived CMs, cardiac fibroblasts (CFs), and cardiac endothelial cells also enhance maturation in easily constructed, scaffold-free, three-dimensional microtissues (MTs). hiPSC-CMs in MTs with CFs showed improved sarcomeric structures with T-tubules, enhanced contractility, and mitochondrial respiration and were electrophysiologically more mature than MTs without CFs. Interactions mediating maturation included coupling between hiPSC-CMs and CFs through connexin 43 (CX43) gap junctions and increased intracellular cyclic AMP (cAMP). Scaled production of thousands of hiPSC-MTs was highly reproducible across lines and differentiated cell batches. MTs containing healthy-control hiPSC-CMs but hiPSC-CFs from patients with arrhythmogenic cardiomyopathy strikingly recapitulated features of the disease. Our MT model is thus a simple and versatile platform for modeling multicellular cardiac diseases that will facilitate industry and academic engagement in high-throughput molecular screening.
