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

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• 作者：Yau-Chi Chan (1)
  Sherwin Ting (2)
  Yee-Ki Lee (1)
  Kwong-Man Ng (1)
  Jiao Zhang (1)
  Zi Chen (1)
  Chung-Wah Siu (1) (3)
  Steve K. W. Oh (2) (4)
  Hung-Fat Tse (1) (3)
  
• 关键词：Human embryonic stem cells ; Cardiomyocytes ; Electrical stimulation
• 刊名：Journal of Cardiovascular Translational Research
• 出版年：2013
• 出版时间：December 2013
• 年：2013
• 卷：6
• 期：6
• 页码：989-999
• 全文大小：
• 作者单位：Yau-Chi Chan (1)
  Sherwin Ting (2)
  Yee-Ki Lee (1)
  Kwong-Man Ng (1)
  Jiao Zhang (1)
  Zi Chen (1)
  Chung-Wah Siu (1) (3)
  Steve K. W. Oh (2) (4)
  Hung-Fat Tse (1) (3)
  
  1. Cardiology Division, Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Hong Kong, Hong Kong SAR, China
  2. Bioprocessing Technology Institute, A*STAR (Agency for Science, Technology and Research), Singapore, Singapore
  3. Research Centre of Heart, Brain, Hormone and Healthy Ageing, The University of Hong Kong, Hong Kong, Hong Kong SAR, China
  4. Stem Cell Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Centros Level 4, 20 Biopolis Way, Singapore, 138668, Singapore
  
• ISSN：1937-5395

文摘

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.