title: Human cardiomyocyte progenitor cell-derived cardiomyocytes display a maturated electrical phenotype.
abstract: Cardiomyocyte progenitor cells (CMPCs) can be isolated from the human heart and differentiated into cardiomyocytes in vitro. A comprehensive assessment of their electrical phenotype upon differentiation is essential to predict potential future applications of this cell source. CMPCs isolated from human fetal heart were differentiated in vitro and examined using immunohistochemistry, Western blotting, RT-PCR, voltage clamp and current clamp techniques. Differentiated cultures presented up to 95% alpha-actinin positive cardiomyocytes. Adherens junction and desmosomal proteins beta-catenin, N-cadherin, desmin and plakophilin2 were upregulated. Expression levels of cardiac connexins were not affected by differentiation, however Cx43 phosphorylation was increased upon differentiation, accompanied by translocation of connexins to the cell border. RT-PCR analysis demonstrated upregulation of all major cardiac ion channel constituents during differentiation. Patch clamp experiments showed that cardiomyocytes had a stable resting membrane potential of -73.4+/-1.8 mV. Infusion of 1 mM BaCl(2) resulted in depolarization to -59.9+/-2.8 mV, indicating I(K1) channel activity. Subsequent voltage clamp experiments confirmed presence of near mature I(Na), I(Ca,L) and I(K1) current densities. Infusion of the I(Kr) blocker Almokalant caused prolongation of the action potential by 40%. Differentiated monolayers were not spontaneously contracting in the absence of serum, but responded to field stimulation, displaying adult ventricular-like action potentials. Human fetal CMPC-derived cardiomyocytes have a homogenous and rather mature electrical phenotype that benefits to in vitro physiology and pharmacology. In the context of cardiac repair, their properties may translate into a reduced pro-arrhythmic risk and enhanced electrical integration upon transplantation.
authors.affiliation: Department of Medical Physiology, Division of Heart and Lungs, University Medical Center Utrecht, Yalelaan 50, 3584 CM Utrecht, The Netherlands.
authors.forename: Teun P
authors.initials: TP
authors.lastname: de Boer
authors.forename: Toon A B
authors.initials: TA
authors.lastname: van Veen
authors.forename: Malin K B
authors.initials: MK
authors.lastname: Jonsson
authors.forename: Bart G J M
authors.initials: BG
authors.lastname: Kok
authors.forename: Corina H G
authors.initials: CH
authors.lastname: Metz
authors.forename: Joost P G
authors.initials: JP
authors.lastname: Sluijter
authors.forename: Pieter A
authors.initials: PA
authors.lastname: Doevendans
authors.forename: Jacques M T
authors.initials: JM
authors.lastname: de Bakker
authors.forename: Marie-José
authors.initials: MJ
authors.lastname: Goumans
authors.forename: Marcel A G
authors.initials: MA
authors.lastname: van der Heyden
mesh_terms: D000889:Anti-Arrhythmia Agents / Q000494:pharmacology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D004594:Electrophysiology; D006321:Heart / Q000187:drug effects / Q000502:physiology; D006801:Humans; D007473:Ion Channels / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D024661:Potassium Channels, Inwardly Rectifying / Q000378:metabolism; D011412:Propanolamines / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology*
chemical_list: D000889:Anti-Arrhythmia Agents; D007473:Ion Channels; D024661:Potassium Channels, Inwardly Rectifying; D011412:Propanolamines; C074334:almokalant