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Suppression of P-cadherin expression as a key regulatory element for embryonic stem cell stemness
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- Takeda Yuka
- Department of Biomedical Sciences, Graduate School of Science and Technology, Kwansei Gakuin University
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- Matsuguchi Shuji
- Department of Biomedical Sciences, Graduate School of Science and Technology, Kwansei Gakuin University
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- Nozaki Sae
- Department of Biomedical Sciences, Graduate School of Science and Technology, Kwansei Gakuin University
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- Mihara Taisei
- Department of Biomedical Sciences, Graduate School of Science and Technology, Kwansei Gakuin University
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- Abe Junya
- Department of Biomedical Sciences, Graduate School of Science and Technology, Kwansei Gakuin University
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- Hirai Yohei
- Department of Biomedical Sciences, Graduate School of Science and Technology, Kwansei Gakuin University
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Description
<p>In embryonic stem (ES) cell colonies, a small subpopulation that changes cell shape and loses pluripotency often appears in two-dimensional (2D) cultures, even in the presence of a stemness factor. We have previously shown that membrane translocation of the syntaxin4, t-SNARE protein contributes to this phenomenon. Here, we show that ES cells in three-dimensional (3D) aggregates do not succumb to extruded syntaxin4 owing to suppressed expression of P-cadherin protein. While extracellular expression of syntaxin4 led to the striking upregulation of P-cadherin mRNA in both 2D and 3D-ES cells, morphological changes and appreciable expression of P-cadherin protein were detected only in 2D-ES cells. Importantly, the introduction of an expression cassette for P-cadherin practically reproduced the effects induced by extracellular syntaxin4, where the transgene product was clearly detected in 2D-, but not 3D-ES cells. An expression construct for P-cadherin-Venus harboring an in-frame insertion of the P2A sequence at the joint region gave fluorescent signals only in the cytoplasm of 2D-ES cells, demonstrating translational regulation of P-cadherin. These results provide the mechanistic insight into the uncontrollable differentiation in 2D-ES cells and shed light on the validity of the “embryoid body protocol commonly used for ES cell handling” for directional differentiation.</p><p>Key words: differentiation, embryoid body, ES cells, P-cadherin, syntaxin4</p>
Journal
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- Cell Structure and Function
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Cell Structure and Function 48 (1), 49-57, 2023
Japan Society for Cell Biology