title: Thymosinβ4 Improves Differentiation and Vascularization of EHTs.
abstract: Induced pluripotent stem cells (iPSC) constitute a powerful tool to study cardiac physiology and represents a promising treatment strategy to tackle cardiac disease. However, iPSCs remain relatively immature after differentiation. Additionally, engineered heart tissue (EHT) has been investigated as a therapy option in preclinical disease models with promising results, although their vascularization and functionality leave room for improvement. Thymosinβ4 (Tβ4) has been shown to promote the differentiation of progenitor cell lines to cardiomyocytes while it also induces angiogenic sprouting and vascular maturation. We examined the potential impact of Tβ4 to enhance maturation of cardiomyocytes from iPSCs. Assessing the expression of transcription factors associated with cardiac differentiation, we were able to demonstrate the increased generation of cells displaying cardiomyocyte characteristics in vitro. Furthermore, we demonstrated, in a zebrafish model of embryonic vascular development, that Tβ4 is crucial for the proper execution of lymphatic and angiogenic vessel sprouting. Finally, utilizing Tβ4-transduced EHTs generated from mice genetically engineered to label endothelial cells in vitro, we show that treatment with Tβ4 promotes vascularization and contractility in EHTs, highlighting Tβ4 as a growth factor improving the formation of cardiomyocytes from iPSC and enhancing the performance of EHTs generated from neonatal cardiomyocytes.
authors.affiliation: I. Medizinische Klinik und Poliklinik, Klinikum rechts der Isar, Munich, Germany; German Center for Cardiovascular Research (DZHK), Partner Site Munich Heart Alliance, Munich, Germany.
authors.forename: Tilman
authors.initials: T
authors.lastname: Ziegler
authors.affiliation: I. Medizinische Klinik und Poliklinik, Klinikum rechts der Isar, Munich, Germany; German Center for Cardiovascular Research (DZHK), Partner Site Munich Heart Alliance, Munich, Germany; Institute for Cardiovascular Prevention, Ludwig Maximilians University, Munich, Germany.
authors.forename: Rabea
authors.initials: R
authors.lastname: Hinkel
authors.affiliation: Institut für Experimentelle Pharmakologie und Toxikologie, Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Stöhr
authors.affiliation: Institut für Experimentelle Pharmakologie und Toxikologie, Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Eschenhagen
authors.affiliation: I. Medizinische Klinik und Poliklinik, Klinikum rechts der Isar, Munich, Germany.
authors.forename: Karl-Ludwig
authors.initials: KL
authors.lastname: Laugwitz
authors.affiliation: Karlsruher Institut für Technologie, Karlsruhe, Germany.
authors.forename: Ferdinand
authors.initials: F
authors.lastname: le Noble
authors.affiliation: Department of Cardiac Surgery, Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), University of Rostock, Rostock, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: David
authors.affiliation: Institut für Experimentelle Pharmakologie und Toxikologie, Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany; German Center for Cardiovascular Research (DZHK), Partner Site Hamburg/Kiel/Lübeck, Hamburg, Germany.
authors.forename: Arne
authors.initials: A
authors.lastname: Hansen
authors.affiliation: I. Medizinische Klinik und Poliklinik, Klinikum rechts der Isar, Munich, Germany; German Center for Cardiovascular Research (DZHK), Partner Site Munich Heart Alliance, Munich, Germany.
authors.forename: Christian
authors.identifier: 0000-0002-3611-1218
authors.initials: C
authors.lastname: Kupatt