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In vitro differentiation of W8B2⁺ human cardiac stem cells: gene expression of ionic channels and spontaneous calcium activity.

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 differentiation of these stem cells has been reported.

Methods: The objectives of this study were thus to analyze (using the TaqMan Low-Density 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 addition, we reveal spontaneous calcium activity or a calcium clock whose kinetics change during the differentiation process. A pharmacological study carried out on differentiated 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 process of stem cells expressing the W8B2 marker.

SUBMITTER: Ayad O

PROVIDER: S-EPMC7646077 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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In vitro differentiation of W8B2<sup>+</sup> human cardiac stem cells: gene expression of ionic channels and spontaneous calcium activity.

Ayad Oualid O Al Sayed Zeina R ZR Sebille Stéphane S Magaud Christophe C Chapotte-Baldacci Charles-Albert CA Jayle Christophe C Faivre Jean-François JF Gaborit Nathalie N Chatelier Aurélien A Bois Patrick P

Cellular & molecular biology letters 20201105 1

<h4>Background</h4>Human cardiac stem cells expressing the W8B2 marker (W8B2<sup>+</sup> 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 differentiation of these stem cells has been reported.<h4>Methods</h4>The objectives of this study were thus to analyze (using the TaqMan Low-Density Array te ...[more]

PMID: 33292162

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Project description:BackgroundRecent studies suggest that ovarian germ line stem cells replenish oocyte-pool in adult stage, and challenge the central doctrine of 'fixed germ cell pool' in mammalian reproductive biology. Two distinct populations of spherical stem cells with high nucleo-cytoplasmic ratio have been recently identified in the adult mammalian ovary surface epithelium (OSE) including nuclear OCT-4A positive very small embryonic-like (VSELs) and cytoplasmic OCT-4 expressing ovarian germ stem cells (OGSCs). Three weeks culture of scraped OSE cells results in spontaneous differentiation of the stem cells into oocyte-like, parthenote-like, embryoid body-like structures and also embryonic stem cell-like colonies whereas epithelial cells attach and transform into a bed of mesenchymal cells. Present study was undertaken, to further characterize ovarian stem cells and to comprehend better the process of spontaneous differentiation of ovarian stem cells into oocyte-like structures in vitro.MethodsOvarian stem cells were enriched by immunomagnetic sorting using SSEA-4 as a cell surface marker and were further characterized. Stem cells and clusters of OGSCs (reminiscent of germ cell nests in fetal ovaries), were characterized by immuno-localization for stem and germ cell specific markers and spontaneous differentiation in OSE cultures was studied by live cell imaging.ResultsDifferential expression of markers specific for pluripotent VSELs (nuclear OCT-4A, SSEA-4, CD133), OGSCs (cytoplasmic OCT-4) primordial germ cells (FRAGILIS, STELLA, VASA) and germ cells (DAZL, GDF-9, SCP-3) were studied. Within one week of culture, stem cells became bigger in size, developed abundant cytoplasm, differentiated into germ cells, revealed presence of Balbiani body-like structure (mitochondrial cloud) and exhibited characteristic cytoplasmic streaming.ConclusionsPresence of germ cell nests, Balbiani body-like structures and cytoplasmic streaming extensively described during fetal ovary development, are indeed well recapitulated during in vitro oogenesis in adult OSE cultures along with characteristic expression of stem/germ cell/oocyte markers. Further studies are required to assess the genetic integrity of in vitro derived oocytes before harnessing their clinical potential. Advance in our knowledge about germ cell differentiation from stem cells will enable researchers to design better in vitro strategies which in turn may have relevance to reproductive biology and regenerative medicine.

Dataset Information

In vitro differentiation of W8B2⁺ human cardiac stem cells: gene expression of ionic channels and spontaneous calcium activity.

Publications

In vitro differentiation of W8B2<sup>+</sup> human cardiac stem cells: gene expression of ionic channels and spontaneous calcium activity.

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Dataset Information

In vitro differentiation of W8B2+ human cardiac stem cells: gene expression of ionic channels and spontaneous calcium activity.

Publications

In vitro differentiation of W8B2<sup>+</sup> human cardiac stem cells: gene expression of ionic channels and spontaneous calcium activity.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

In vitro differentiation of W8B2⁺ human cardiac stem cells: gene expression of ionic channels and spontaneous calcium activity.