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Uzun AU, Mannhardt I, Breckwoldt K, et al. Ca(2+)-Currents in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Effects of Two Different Culture Conditions. Front Pharmacol. 2016;7:300doi: 10.3389/fphar.2016.00300.
Uzun, A. U., Mannhardt, I., Breckwoldt, K., Horváth, A., Johannsen, S. S., Hansen, A., Eschenhagen, T., & Christ, T. (2016). Ca(2+)-Currents in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Effects of Two Different Culture Conditions. Frontiers in pharmacology, 7300. https://doi.org/10.3389/fphar.2016.00300
Uzun, Ahmet U, et al. "Ca(2+)-Currents in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Effects of Two Different Culture Conditions." Frontiers in pharmacology vol. 7 (2016): 300. doi: https://doi.org/10.3389/fphar.2016.00300
Uzun AU, Mannhardt I, Breckwoldt K, Horváth A, Johannsen SS, Hansen A, Eschenhagen T, Christ T. Ca(2+)-Currents in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Effects of Two Different Culture Conditions. Front Pharmacol. 2016 Sep 12;7:300. doi: 10.3389/fphar.2016.00300. eCollection 2016. PMID: 27672365; PMCID: PMC5018497.
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Front Pharmacol. 2016 Sep 12;7:300. doi: 10.3389/fphar.2016.00300. eCollection 2016.

Ca(2+)-Currents in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Effects of Two Different Culture Conditions.

Frontiers in pharmacology

Ahmet U Uzun, Ingra Mannhardt, Kaja Breckwoldt, András Horváth, Silke S Johannsen, Arne Hansen, Thomas Eschenhagen, Torsten Christ

Affiliations

  1. Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-EppendorfHamburg, Germany; Partner Site Hamburg/Kiel/Lübeck, German Centre for Cardiovascular Research (DZHK)Hamburg, Germany.
  2. Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-EppendorfHamburg, Germany; Partner Site Hamburg/Kiel/Lübeck, German Centre for Cardiovascular Research (DZHK)Hamburg, Germany; Department of Pharmacology and Pharmacotherapy, Faculty of Medicine, University of SzegedSzeged, Hungary.
  3. Partner Site Hamburg/Kiel/Lübeck, German Centre for Cardiovascular Research (DZHK)Hamburg, Germany; Department of General and Interventional Cardiology, University Heart Center HamburgHamburg, Germany.

PMID: 27672365 PMCID: PMC5018497 DOI: 10.3389/fphar.2016.00300

Abstract

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) provide a unique opportunity to study human heart physiology and pharmacology and repair injured hearts. The suitability of hiPSC-CM critically depends on how closely they share physiological properties of human adult cardiomyocytes (CM). Here we investigated whether a 3D engineered heart tissue (EHT) culture format favors maturation and addressed the L-type Ca(2+)-current (ICa,L) as a readout. The results were compared with hiPSC-CM cultured in conventional monolayer (ML) and to our previous data from human adult atrial and ventricular CM obtained when identical patch-clamp protocols were used. HiPSC-CM were two- to three-fold smaller than adult CM, independently of culture format [capacitance ML 45 ± 1 pF (n = 289), EHT 45 ± 1 pF (n = 460), atrial CM 87 ± 3 pF (n = 196), ventricular CM 126 ± 8 pF (n = 50)]. Only 88% of ML cells showed ICa, but all EHT. Basal ICa density was 10 ± 1 pA/pF (n = 207) for ML and 12 ± 1 pA/pF (n = 361) for EHT and was larger than in adult CM [7 ± 1 pA/pF (p < 0.05, n = 196) for atrial CM and 6 ± 1 pA/pF (p < 0.05, n = 47) for ventricular CM]. However, ML and EHT showed robust T-type Ca(2+)-currents (ICa,T). While (-)-Bay K 8644, that activates ICa,L directly, increased ICa,Lto the same extent in ML and EHT, β1- and β2-adrenoceptor effects were marginal in ML, but of same size as (-)-Bay K 8644 in EHT. The opposite was true for serotonin receptors. Sensitivity to β1 and β2-adrenoceptor stimulation was the same in EHT as in adult CM (-logEC50: 5.9 and 6.1 for norepinephrine (NE) and epinephrine (Epi), respectively), but very low concentrations of Rp-8-Br-cAMPS were sufficient to suppress effects (-logEC50: 5.3 and 5.3 respectively for NE and Epi). Taken together, hiPSC-CM express ICa,L at the same density as human adult CM, but, in contrast, possess robust ICa,T. Increased effects of catecholamines in EHT suggest more efficient maturation.

Keywords: 5-hydroxytryptamine; L-type Ca2+-channel; T-type Ca2+-channel; human induced pluripotent stem cell-derived cardiomyocytes; protein kinase A; β-adrenoceptor

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