title: Three-dimensional cardiac microtissues composed of cardiomyocytes and endothelial cells co-differentiated from human pluripotent stem cells.
abstract: Cardiomyocytes and endothelial cells in the heart are in close proximity and in constant dialogue. Endothelium regulates the size of the heart, supplies oxygen to the myocardium and secretes factors that support cardiomyocyte function. Robust and predictive cardiac disease models that faithfully recapitulate native human physiologyin vitrowould therefore ideally incorporate this cardiomyocyte-endothelium crosstalk. Here, we have generated and characterized human cardiac microtissuesin vitrothat integrate both cell types in complex 3D structures. We established conditions for simultaneous differentiation of cardiomyocytes and endothelial cells from human pluripotent stem cells following initial cardiac mesoderm induction. The endothelial cells expressed cardiac markers that were also present in primary cardiac microvasculature, suggesting cardiac endothelium identity. These cell populations were further enriched based on surface markers expression, then recombined allowing development of beating 3D structures termed cardiac microtissues. Thisin vitromodel was robustly reproducible in both embryonic and induced pluripotent stem cells. It thus represents an advanced human stem cell-based platform for cardiovascular disease modelling and testing of relevant drugs.
authors.affiliation: Department of Anatomy and Embryology, Leiden University Medical Center, Leiden 2333ZC, The Netherlands.
authors.forename: Elisa
authors.identifier: 0000-0001-9233-235X
authors.initials: E
authors.lastname: Giacomelli
authors.affiliation: Department of Anatomy and Embryology, Leiden University Medical Center, Leiden 2333ZC, The Netherlands.
authors.forename: Milena
authors.identifier: 0000-0001-5380-6743
authors.initials: M
authors.lastname: Bellin
authors.affiliation: Department of Anatomy and Embryology, Leiden University Medical Center, Leiden 2333ZC, The Netherlands.
authors.forename: Luca
authors.identifier: 0000-0002-4129-6632
authors.initials: L
authors.lastname: Sala
authors.affiliation: Department of Anatomy and Embryology, Leiden University Medical Center, Leiden 2333ZC, The Netherlands.
authors.forename: Berend J
authors.initials: BJ
authors.lastname: van Meer
authors.affiliation: Department of Anatomy and Embryology, Leiden University Medical Center, Leiden 2333ZC, The Netherlands.
authors.forename: Leon G J
authors.initials: LG
authors.lastname: Tertoolen
authors.affiliation: Department of Anatomy and Embryology, Leiden University Medical Center, Leiden 2333ZC, The Netherlands.
authors.forename: Valeria V
authors.identifier: 0000-0002-1169-2802
authors.initials: VV
authors.lastname: Orlova
authors.affiliation: Department of Anatomy and Embryology, Leiden University Medical Center, Leiden 2333ZC, The Netherlands c.l.mummery@lumc.nl.|Department of Applied Stem Cell Technologies, University of Twente, Enschede 7500AE, The Netherlands.
authors.forename: Christine L
authors.identifier: 0000-0002-4549-6535
authors.initials: CL
authors.lastname: Mummery
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism; D002454:Cell Differentiation; D002469:Cell Separation; D055724:Electrophysiological Phenomena; D042783:Endothelial Cells / Q000166:cytology*; D006801:Humans; D008648:Mesoderm / Q000166:cytology; D032383:Myocytes, Cardiac / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D012518:Sarcomeres / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
chemical_list: D018952:Antigens, CD34; D019010:Vascular Cell Adhesion Molecule-1
keywords: Cardiac microtissue (MT); Human pluripotent stem cell-derived cardiomyocytes; Human pluripotent stem cell-derived endothelial cells; Mesoderm induction; Three-dimensional culture model