title: Myocardial commitment from human pluripotent stem cells: Rapid production of human heart grafts.
abstract: Genome editing on human pluripotent stem cells (hPSCs) together with the development of protocols for organ decellularization opens the door to the generation of autologous bioartificial hearts. Here we sought to generate for the first time a fluorescent reporter human embryonic stem cell (hESC) line by means of Transcription activator-like effector nucleases (TALENs) to efficiently produce cardiomyocyte-like cells (CLCs) from hPSCs and repopulate decellularized human heart ventricles for heart engineering. In our hands, targeting myosin heavy chain locus (MYH6) with mCherry fluorescent reporter by TALEN technology in hESCs did not alter major pluripotent-related features, and allowed for the definition of a robust protocol for CLCs production also from human induced pluripotent stem cells (hiPSCs) in 14 days. hPSCs-derived CLCs (hPSCs-CLCs) were next used to recellularize acellular cardiac scaffolds. Electrophysiological responses encountered when hPSCs-CLCs were cultured on ventricular decellularized extracellular matrix (vdECM) correlated with significant increases in the levels of expression of different ion channels determinant for calcium homeostasis and heart contractile function. Overall, the approach described here allows for the rapid generation of human cardiac grafts from hPSCs, in a total of 24 days, providing a suitable platform for cardiac engineering and disease modeling in the human setting.
authors.affiliation: Pluripotent Stem Cells and Activation of Endogenous Tissue Programs for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Garreta
authors.affiliation: Pluripotent Stem Cells and Activation of Endogenous Tissue Programs for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain; Center of Regenerative Medicine in Barcelona (CMRB), Barcelona, Spain.
authors.forename: Lorena
authors.initials: L
authors.lastname: de Oñate
authors.affiliation: Department of Cardiology, Hospital General Universitario Gregorio Marañón, Universidad Complutense de Madrid, Spain; Cell Production Unit, Department of Cardiology, Instituto de Investigación Sanitaria Hospital Gregorio Marañón (IiSGM), Madrid, Spain.
authors.forename: M Eugenia
authors.initials: ME
authors.lastname: Fernández-Santos
authors.affiliation: Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain; University of Barcelona, Barcelona, Spain.
authors.forename: Roger
authors.initials: R
authors.lastname: Oria
authors.affiliation: Pluripotent Stem Cells and Activation of Endogenous Tissue Programs for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain.
authors.forename: Carolina
authors.initials: C
authors.lastname: Tarantino
authors.affiliation: Bioartifical Organs Laboratory, Instituto de Investigación Sanitaria Hospital Gregorio Marañón (IiSGM), Madrid, Spain.
authors.forename: Andreu M
authors.initials: AM
authors.lastname: Climent
authors.affiliation: Pluripotent Stem Cells and Activation of Endogenous Tissue Programs for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain.
authors.forename: Andrés
authors.initials: A
authors.lastname: Marco
authors.affiliation: Pluripotent Stem Cells and Activation of Endogenous Tissue Programs for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain.
authors.forename: Mireia
authors.initials: M
authors.lastname: Samitier
authors.affiliation: Biomimetic Systems for Cell Engineering, Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Martínez
authors.affiliation: Biomimetic Systems for Cell Engineering, Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain.
authors.forename: Maria
authors.initials: M
authors.lastname: Valls-Margarit
authors.affiliation: National Transplant Organization (ONT), Spanish Ministry of Health and Consumption, Spain.
authors.forename: Rafael
authors.initials: R
authors.lastname: Matesanz
authors.affiliation: Center for Cardiovascular Repair, University of Minnesota, Minneapolis, MN, USA; Department of Regenerative Medicine Research, Texas Heart Institute, Houston, TX, USA; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Doris A
authors.initials: DA
authors.lastname: Taylor
authors.affiliation: Department of Cardiology, Hospital General Universitario Gregorio Marañón, Universidad Complutense de Madrid, Spain; Cell Production Unit, Department of Cardiology, Instituto de Investigación Sanitaria Hospital Gregorio Marañón (IiSGM), Madrid, Spain. Electronic address: faviles@secardiologia.es.
authors.forename: Francisco
authors.initials: F
authors.lastname: Fernández-Avilés
authors.affiliation: Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, CA, USA. Electronic address: belmonte@salk.edu.
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Izpisua Belmonte
authors.affiliation: Pluripotent Stem Cells and Activation of Endogenous Tissue Programs for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain. Electronic address: nmontserrat@ibecbarcelona.eu.
authors.forename: Nuria
authors.initials: N
authors.lastname: Montserrat
mesh_terms: D024722:Cardiac Myosins / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D055724:Electrophysiological Phenomena / Q000187:drug effects; D005109:Extracellular Matrix / Q000378:metabolism; D056426:Genetic Loci; D016027:Heart Transplantation; D006352:Heart Ventricles / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D009206:Myocardium / Q000166:cytology*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011509:Proteoglycans / Q000494:pharmacology; D000069896:Transcription Activator-Like Effector Nucleases
chemical_list: D004338:Drug Combinations; D007797:Laminin; C094239:MYH6 protein, human; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D000069896:Transcription Activator-Like Effector Nucleases; D024722:Cardiac Myosins; D018995:Myosin Heavy Chains
keywords: Cardiac function; Extracellular matrix; Gene targeting; Pluripotent stem cells