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Human Cell

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Enhancement of connexin30.3 expression in mouse embryonic stem cell line EB3 in response to cell–cell contacts

  • Naruwa Tokunaga
  • Ryota Kishi
  • Tomoko Sasai
  • Mikako SaitoEmail author
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Abstract

To clarify the potential role of gap junction in cell–cell contact response, the expression of connexin30.3 gene (Cx30.3), a specifically expressed isoform in undifferentiated state of mouse embryonic stem (ES) cell line EB3 was investigated under different cell–cell contact conditions. ES cells were cultured by hanging drop culture method to increase cell–cell contact frequency. As control, a single cell culture was conducted. After culture for 12 h, the Cx30.3 expression level in hanging drop culture reached 1.73-fold that of the control (p < 0.001). By contrast, connexin43 gene (Cx43), a ubiquitously expressed gene, showed no difference between both cultures. The experiment of E-cadherin inhibition and β-catenin knockdown suggested the action of E-cadherin upstream of the Cx30.3 regulating pathway. The cell–cell contacts with different cell lines such as HeLa cells and B16/BL6 caused no effect on the Cx30.3 in ES cells. These suggest a potential role of Cx30.3 as a cell–cell contact signal mediator partially regulated by E-cadherin signaling.

Keywords

Embryonic stem cell Connexin30.3 Cell–cell contact Hanging drop culture Cadherin signaling 

Notes

Acknowledgements

This work was partially supported by a Grant-in-Aid for Scientific Research (C) Grant number JP16K07343.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Japan Human Cell Society and Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  • Naruwa Tokunaga
    • 1
  • Ryota Kishi
    • 1
  • Tomoko Sasai
    • 1
  • Mikako Saito
    • 1
    Email author
  1. 1.Department of Biotechnology and Life ScienceTokyo University of Agriculture and TechnologyKoganeiJapan

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