Gap Junctional Coupling with Cardiomyocytes is Necessary but Not Sufficient for Cardiomyogenic Differentiation of Cocultured Human Mesenchymal Stem Cells§

Authors

  • Arti A. Ramkisoensing,

    1. Department of Cardiology,Laboratory of Experimental Cardiology, Leiden University Medical Center, Leiden, The Netherlands
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  • DaniëL A. Pijnappels,

    1. Department of Cardiology,Laboratory of Experimental Cardiology, Leiden University Medical Center, Leiden, The Netherlands
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  • Jim Swildens,

    1. Department of Molecular Cell Biology,Laboratory of Experimental Cardiology, Leiden University Medical Center, Leiden, The Netherlands
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  • Marie José Goumans,

    1. Department of Molecular Cell Biology,Laboratory of Experimental Cardiology, Leiden University Medical Center, Leiden, The Netherlands
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  • Willem E. Fibbe,

    1. Laboratory of Experimental Cardiology, Department of Hematology, Leiden University Medical Center, Leiden, The Netherlands
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  • Martin J. Schalij,

    1. Department of Cardiology,Laboratory of Experimental Cardiology, Leiden University Medical Center, Leiden, The Netherlands
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  • Antoine A.F. de Vries,

    Corresponding author
    1. Department of Cardiology,Laboratory of Experimental Cardiology, Leiden University Medical Center, Leiden, The Netherlands
    2. Department of Molecular Cell Biology,Laboratory of Experimental Cardiology, Leiden University Medical Center, Leiden, The Netherlands
    • Department of Cardiology, Leiden University Medical Center, Leiden 2300 RC, The Netherlands
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    • Telephone: +31-715262020; Fax: +31-715266809

    • contributed equally to this article.

  • Douwe E. Atsma

    Corresponding author
    1. Department of Cardiology,Laboratory of Experimental Cardiology, Leiden University Medical Center, Leiden, The Netherlands
    • Department of Cardiology, Leiden University Medical Center, Leiden 2300 RC, The Netherlands
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    • contributed equally to this article.

    • Telephone: +31-715262020; Fax: +31-715266809


  • Author Contributions: A.A.R. and D.A.P.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript; J.S.: provision of study material, data analysis and interpretation, manuscript writing, and final approval of manuscript; M.J.G. and W.E.F.: provision of study material, administrative support, financial support, and final approval of manuscript; M.J.S.: financial support, data analysis and interpretation, manuscript writing, and final approval of manuscript; A.A.F.V.: conception and design, data analysis and interpretation, manuscript writing, and final approval of manuscript; D.E.A.: conception and design, financial support, data analysis and interpretation, manuscript writing, and final approval of manuscript.

  • Disclosure of potential conflicts of interest is found at the end of this article.

  • §

    First published online in STEM CELLSEXPRESS Month 00, 2012.

Abstract

Gap junctional coupling is important for functional integration of transplanted cells with host myocardium. However, the role of gap junctions in cardiomyogenic differentiation of transplanted cells has not been directly investigated. The objective of this work is to study the role of connexin43 (Cx43) in cardiomyogenic differentiation of human mesenchymal stem cells (hMSCs). Knockdown of Cx43 gene expression (Cx43↓) was established in naturally Cx43-rich fetal amniotic membrane (AM) hMSCs, while Cx43 was overexpressed (Cx43↑) in inherently Cx43-poor adult adipose tissue (AT) hMSCs. The hMSCs were exposed to cardiomyogenic stimuli by coincubation with neonatal rat ventricular cardiomyocytes (nrCMCs) for 10 days. Differentiation was assessed by immunostaining and whole-cell current clamping. To establish whether the effects of Cx43 knockdown could be rescued, Cx45 was overexpressed in Cx43↓ fetal AM hMSCs. Ten days after coincubation, not a single Cx43↓ fetal AM hMSC, control adult AT MSC, or Cx43↑ adult AT mesenchymal stem cell (MSC) expressed α-actinin, while control fetal AM hMSCs did (2.2% ± 0.4%, n = 5,000). Moreover, functional cardiomyogenic differentiation, based on action potential recordings, occurred only in control fetal AM hMSCs. Of interest, Cx45 overexpression in Cx43↓ fetal AM hMSCs restored their ability to undergo cardiomyogenesis (1.6% ± 0.4%, n = 2,500) in coculture with nrCMCs. Gap junctional coupling is required for differentiation of fetal AM hMSCs into functional CMCs after coincubation with nrCMCs. Heterocellular gap junctional coupling thus plays an important role in the transfer of cardiomyogenic signals from nrCMCs to fetal hMSCs but is not sufficient to induce cardiomyogenic differentiation in adult AT hMSCs. STEM CELLS2012;30:1236–1245

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