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Electrical Stimulation Promotes Maturation of Cardiomyocytes Derived from Human Embryonic Stem Cells

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Abstract

While human embryonic stem cells (hESCs) can differentiate into functional cardiomyocytes, their immature phenotypes limit their therapeutic application for myocardial regeneration. We sought to determine whether electrical stimulation could enhance the differentiation and maturation of hESC-derived cardiomyocytes. Cardiac differentiation was induced in a HES3 hESC line via embryoid bodies formation treated with a p38 MAP kinase inhibitor. Detailed molecular and functional analysis were performed in those hESC-derived cardiomyocytes cultured for 4 days in the absence or presence of electrical field stimulation (6.6 V/cm, 1 Hz, and 2 ms pulses) using an eight-channel C-Pace stimulator (Ion-Optics Co., MA). Upon electrical stimulation, quantitative polymerase chain reaction demonstrated significant upregulation of cardiac-specific gene expression including HCN1, MLC2V, SCN5A, SERCA, Kv4.3, and GATA4; immunostaining and flow cytometry analysis revealed cellular elongation and an increased proportion of troponin-T positive cells (6.3 ± 1.2 % vs. 15.8 ± 2.1 %; n = 3, P < 0.01). Electrophysiological studies showed an increase in the proportion of ventricular-like hESC-derived cardiomyocytes (48 vs. 29 %, P < 0.05) with lengthening of their action potential duration at 90 % repolarization (387.7 ± 35.35; n = 11 vs. 291.8 ± 20.82; n = 10, P < 0.05) and 50 % repolarization (313.9 ± 27.94; n = 11 vs. 234.0 ± 16.10; n = 10, P < 0.05) after electrical stimulation. Nonetheless, the membrane diastolic potentials and action potential upstrokes of different hESC-derived cardiomyocyte phenotypes, and the overall beating rate remained unchanged (all P > 0.05). Fluorescence confocal imaging revealed that electrical stimulation significantly increased both spontaneous and caffeine-induced calcium flux in the hESC-derived cardiomyocytes (approximately 1.6-fold for both cases; P < 0.01). In conclusion, electrical field stimulation increased the expression of cardiac-specific genes and the yield of differentiation, promoted ventricular-like phenotypes, and improved the calcium handling of hESC-derived cardiomyocytes.

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Acknowledgments

This study was supported by Hong Kong Research Grant Council (HKU 8/CRF/09, HKU 8/CRF/10, HKU 780110 M to H.F.T), Theme-based Research Scheme (T12-705/11 to C.W.S and H.F.T), and CRCG Small Project Funding of University of Hong Kong (Y.C.C); Agency for Science Technology and Research (S.T. and S.K.O).

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The authors have nothing to declare.

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Authors and Affiliations

  1. Cardiology Division, Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Hong Kong, Hong Kong SAR, China

    Yau-Chi Chan, Yee-Ki Lee, Kwong-Man Ng, Jiao Zhang, Zi Chen, Chung-Wah Siu & Hung-Fat Tse

  2. Bioprocessing Technology Institute, A*STAR (Agency for Science, Technology and Research), Singapore, Singapore

    Sherwin Ting & Steve K. W. Oh

  3. Research Centre of Heart, Brain, Hormone and Healthy Ageing, The University of Hong Kong, Hong Kong, Hong Kong SAR, China

    Chung-Wah Siu & Hung-Fat Tse

  4. Stem Cell Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Centros Level 4, 20 Biopolis Way, Singapore, 138668, Singapore

    Steve K. W. Oh

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  1. Yau-Chi Chan
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Correspondence to Hung-Fat Tse.

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Associate Editor Jennifer L. Hall oversaw the review of this article.

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Chan, YC., Ting, S., Lee, YK. et al. Electrical Stimulation Promotes Maturation of Cardiomyocytes Derived from Human Embryonic Stem Cells. J. of Cardiovasc. Trans. Res. 6, 989–999 (2013). https://doi.org/10.1007/s12265-013-9510-z

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  • DOI: https://doi.org/10.1007/s12265-013-9510-z

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