title: The influence of physiological matrix conditions on permanent culture of induced pluripotent stem cell-derived cardiomyocytes.
abstract: Cardiomyocytes (CMs) from induced pluripotent stem (iPS) cells mark an important achievement in the development of in vitro pharmacological, toxicological and developmental assays and in the establishment of protocols for cardiac cell replacement therapy. Using CMs generated from murine embryonic stem cells and iPS cells we found increased cell-matrix interaction and more matured embryoid body (EB) structures in iPS cell-derived EBs. However, neither suspension-culture in form of purified cardiac clusters nor adherence-culture on traditional cell culture plastic allowed for extended culture of CMs. CMs grown for five weeks on polystyrene exhibit signs of massive mechanical stress as indicated by α-smooth muscle actin expression and loss of sarcomere integrity. Hydrogels from polyacrylamide allow adapting of the matrix stiffness to that of cardiac tissue. We were able to eliminate the bottleneck of low cell adhesion using 2,5-Dioxopyrrolidin-1-yl-6-acrylamidohexanoate as a crosslinker to immobilize matrix proteins on the gels surface. Finally we present an easy method to generate polyacrylamide gels with a physiological Young's modulus of 55 kPa and defined surface ligand, facilitating the culture of murine and human iPS-CMs, removing excess mechanical stresses and reducing the risk of tissue culture artifacts exerted by stiff substrates.
authors.affiliation: Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of Cologne, Cologne, Germany.
authors.forename: Carlos O
authors.initials: CO
authors.lastname: Heras-Bautista
authors.affiliation: Fraunhofer Institute for Biomedical Engineering IBMT, St. Ingbert, Germany.
authors.forename: Alisa
authors.initials: A
authors.lastname: Katsen-Globa
authors.affiliation: Department of Chemistry, University of Cologne, Germany.
authors.forename: Nils E
authors.initials: NE
authors.lastname: Schloerer
authors.affiliation: Institute of Complex Systems, ICS-7: Biomechanics, Forschungszentrum Jülich GmbH, Jülich, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Dieluweit
authors.affiliation: Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of Cologne, Cologne, Germany; Department of Pediatrics, Cairo University, Cairo, Egypt; Department of Paediatric Cardiology, University Clinics of Cologne, Cologne, Germany.
authors.forename: Osama M
authors.initials: OM
authors.lastname: Abd El Aziz
authors.affiliation: Department of Internal Medicine III, University Clinics of Cologne, Cologne, Germany.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Peinkofer
authors.affiliation: Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of Cologne, Cologne, Germany; Department of Pediatrics, Cairo University, Cairo, Egypt; Department of Paediatric Cardiology, University Clinics of Cologne, Cologne, Germany.
authors.forename: Wael A
authors.initials: WA
authors.lastname: Attia
authors.affiliation: Department of Paediatric Cardiology, University Clinics of Cologne, Cologne, Germany; Division of Pediatric Cardiology, University Children's Hospital, Giessen, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Khalil
authors.affiliation: Department of Paediatric Cardiology, University Clinics of Cologne, Cologne, Germany.
authors.forename: Konrad
authors.initials: K
authors.lastname: Brockmeier
authors.affiliation: Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of Cologne, Cologne, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hescheler
authors.affiliation: Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of Cologne, Cologne, Germany; Department of Paediatric Cardiology, University Clinics of Cologne, Cologne, Germany. Electronic address: kurt.pfannkuche@uni-koeln.de.
authors.forename: Kurt
authors.initials: K
authors.lastname: Pfannkuche
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003432:Cross-Linking Reagents / Q000737:chemistry*; D055119:Elastic Modulus; D058732:Embryoid Bodies / Q000166:cytology; D016326:Extracellular Matrix Proteins / Q000737:chemistry; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D051379:Mice; D008958:Models, Molecular; D032383:Myocytes, Cardiac / Q000166:cytology*; D012518:Sarcomeres / Q000648:ultrastructure; D000099166:Polyacrylamides
chemical_list: D000180:Acrylic Resins; D001672:Biocompatible Materials; D003432:Cross-Linking Reagents; D016326:Extracellular Matrix Proteins; D020100:Hydrogels; D055767:Immobilized Proteins; D000099166:Polyacrylamides
keywords: Cardiomyocyte; Cell adhesion; Cell culture; Cell viability; Cross-linking; Hydrogel