In vitro differentiation of W8B2 + human cardiac stem cells: gene expression of ionic channels and spontaneous calcium a
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RESEARCH LETTER
Cellular & Molecular Biology Letters
Open Access
In vitro differentiation of W8B2+ human cardiac stem cells: gene expression of ionic channels and spontaneous calcium activity Oualid Ayad1, Zeina R. Al Sayed3, Stéphane Sebille1, Christophe Magaud1, Charles‑Albert Chapotte‑Baldacci1, Christophe Jayle2, Jean‑François Faivre1, Nathalie Gaborit3, Aurélien Chatelier1† and Patrick Bois1*†
*Correspondence: Patrick.Bois@univ‑poitiers.fr † Aurélien Chatelier and Patrick Bois have contributed equally to this work 1 University of Poitiers Signalisation et Transports Ioniques Membranaires, EA7349, Poitiers Cedex 09, France Full list of author information is available at the end of the article
Abstract Background: Human cardiac stem cells expressing the W8B2 marker ( W8B2+ CSCs) were recently identified and proposed as a new model of multipotent CSCs capable of differentiating into smooth muscle cells, endothelial cells and immature myocytes. Nevertheless, no characterization of ion channel or calcium activity during the differen‑ tiation of these stem cells has been reported. Methods: The objectives of this study were thus to analyze (using the TaqMan LowDensity Array technique) the gene profile of W8B2+ CSCs pertaining to the regulation of ion channels, transporters and other players involved in the calcium homeostasis of these cells. We also analyzed spontaneous calcium activity (via the GCaMP calcium probe) during the in vitro differentiation of W8B2+ CSCs into cardiac myocytes. Results: Our results show an entirely different electrophysiological genomic profile between W8B2+ CSCs before and after differentiation. Some specific nodal genes, such as Tbx3, HCN, ICaT, L, KV, and NCX, are overexpressed after this differentiation. In addi‑ tion, we reveal spontaneous calcium activity or a calcium clock whose kinetics change during the differentiation process. A pharmacological study carried out on differenti‑ ated W8B2+ CSCs showed that the NCX exchanger and IP3 stores play a fundamental role in the generation of these calcium oscillations. Conclusions: Taken together, the present results provide important information on ion channel expression and intrinsic calcium dynamics during the differentiation pro‑ cess of stem cells expressing the W8B2 marker. Keywords: W8B2+ human cardiac stem cell, Cardiac differentiation, Calcium activity, Oscillations, Ion channels, GCaMP
Background The existence in healthy and pathological human cardiac tissue of rare populations of multipotent cardiac stem cells (CSCs) that are capable of differentiating into myocytes, smooth muscle cells and endothelial cells has been demonstrated by several groups [1– 4]. Several types of CSCs have been isolated and characterized based on the expression
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