Fetal bovine serum enables cardiac differentiation of human embryonic stem cells

Authors

  • Esther Bettiol,

    1. Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, CMU, 1 rue Michel-Servet, 1211 Geneva 4, Geneva, Switzerland
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  • Laura Sartiani,

    1. Center of Molecular Medicine (C.I.M.M.B.A.), University of Florence, Firenze, Italy
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  • Laurie Chicha,

    1. Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, CMU, 1 rue Michel-Servet, 1211 Geneva 4, Geneva, Switzerland
      Tel: +41 22 379 5257
      Fax: +41 22 379 5479
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  • Karl Heinz Krause,

    1. Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, CMU, 1 rue Michel-Servet, 1211 Geneva 4, Geneva, Switzerland
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      Fax: +41 22 379 5479
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  • Elisabetta Cerbai,

    1. Center of Molecular Medicine (C.I.M.M.B.A.), University of Florence, Firenze, Italy
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  • Marisa E. Jaconi

    Corresponding author
    1. Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, CMU, 1 rue Michel-Servet, 1211 Geneva 4, Geneva, Switzerland
      Tel: +41 22 379 5257
      Fax: +41 22 379 5479
      ✉ E-mail: Marisa.Jaconi@medecine.unige.ch
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✉ E-mail: Marisa.Jaconi@medecine.unige.ch

Abstract

Abstract During development, cardiac commitment within the mesoderm requires endoderm-secreted factors. Differentiation of embryonic stem cells into the three germ layers in vitro recapitulates developmental processes and can be influenced by supplements added to culture medium. Hence, we investigated the effect of fetal bovine serum (FBS) and KnockOut™ serum replacement (SR) on germ layers specification and cardiac differentiation of H1 human embryonic stem cells (hESC) within embryoid bodies (EB). At the time of EB formation, FBS triggered an increased apoptosis. As assessed by quantitative PCR on 4-, 10-, and 20-day-old EB, FBS promoted a faster down-regulation of pluripotency marker Oct4 and an increased expression of endodermal (Sox17, α-fetoprotein, AFP) and mesodermal genes (Brachyury, CSX). While neuronal and hematopoietic differentiation occurred in both supplements, spontaneously beating cardiomyocytes were only observed in FBS. Action potential (AP) morphology of hESC-derived cardiomyocytes indicated that ventricular cells were present only after 2 months of culture. However, quantification of myosin light chain 2 ventricular (mlc2v)-positive areas revealed that mlc2v-expressing cardiomyocytes could be detected already after 2 weeks of differentiation, but not in all beating clusters. In conclusion, FBS enabled cardiac differentiation of hESC, likely in an endodermal-dependent pathway. Among cardiac cells, ventricular cardiomyocytes differentiated over time, but not as the predominant cardiac cell subtype.

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