Differential Sarcomere and Electrophysiological Maturation of Human iPSC-Derived Cardiac Myocytes in Monolayer vs. Aggregation-Based Differentiation Protocols - Sorbonne Université
Article Dans Une Revue International Journal of Molecular Sciences Année : 2017

Differential Sarcomere and Electrophysiological Maturation of Human iPSC-Derived Cardiac Myocytes in Monolayer vs. Aggregation-Based Differentiation Protocols

Dorota Jeziorowska
  • Fonction : Auteur
Vincent Fontaine
Charlène Jouve
  • Fonction : Auteur
Eric Villard
  • Fonction : Auteur
David Akbar
  • Fonction : Auteur
Valérie Letang
  • Fonction : Auteur
Pauline Cervello
  • Fonction : Auteur
Jean-Michiel Itier
  • Fonction : Auteur
Marie-Pierre Pruniaux
  • Fonction : Auteur
Jean-Sébastien Hulot
  • Fonction : Auteur

Résumé

Human induced pluripotent stem cells (iPSCs) represent a powerful human model to study cardiac disease in vitro, notably channelopathies and sarcomeric cardiomyopathies. Different protocols for cardiac differentiation of iPSCs have been proposed either based on embroid body formation (3D) or, more recently, on monolayer culture (2D). We performed a direct comparison of the characteristics of the derived cardiomyocytes (iPSC-CMs) on day 27 ± 2 of differentiation between 3D and 2D differentiation protocols with two different Wnt-inhibitors were compared: IWR1 (inhibitor of Wnt response) or IWP2 (inhibitor of Wnt production). We firstly found that the level of Troponin T (TNNT2) expression measured by FACS was significantly higher for both 2D protocols as compared to the 3D protocol. In the three methods, iPSC-CM show sarcomeric structures. However, iPSC-CM generated in 2D protocols constantly displayed larger sarcomere lengths as compared to the 3D protocol. In addition, mRNA and protein analyses reveal higher cTNi to ssTNi ratios in the 2D protocol using IWP2 as compared to both other protocols, indicating a higher sarcomeric maturation. Differentiation of cardiac myocytes with 2D monolayer-based protocols and the use of IWP2 allows the production of higher yield of cardiac myocytes that have more suitable characteristics to study sarcomeric cardiomyopathies.
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Dates et versions

hal-03937368 , version 1 (13-01-2023)

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Citer

Dorota Jeziorowska, Vincent Fontaine, Charlène Jouve, Eric Villard, Sébastien Dussaud, et al.. Differential Sarcomere and Electrophysiological Maturation of Human iPSC-Derived Cardiac Myocytes in Monolayer vs. Aggregation-Based Differentiation Protocols. International Journal of Molecular Sciences, 2017, 18 (6), pp.1173. ⟨10.1126/scisignal.2004302⟩. ⟨hal-03937368⟩
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