Reference : Epithelial-mesenchymal transition process in human embryonic stem cells cultured in f...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
Epithelial-mesenchymal transition process in human embryonic stem cells cultured in feeder-free conditions.
Ullmann, U. [ > > ]
In'T Veld, P. [ > > ]
Gilles, Christine mailto [Université de Liège - ULiège > Département des sciences cliniques > Labo de biologie des tumeurs et du développement >]
Sermon, K. [ > > ]
De Rycke, M. [ > > ]
Van de Velde, H. [ > > ]
Van Steirteghem, A. [ > > ]
Liebears, I. [ > > ]
Molecular Human Reproduction
Oxford University Press
Yes (verified by ORBi)
United Kingdom
[en] human embryonic stem cells ; feeder-free culture ; matrigel ; differentiation ; epithelial–mesenchymal transition
[en] Feeder-free human embryonic stem cell (hESC) culture is associated with the presence of mesenchymal-like cells appearing at the periphery of the colonies. The aim of this study was to identify this early differentiation process. Long-term feeder-free hESC cultures using matrigel and conditioned medium from mouse and from human origin revealed that the appearance of mesenchymal-like cells was similar regardless of the conditioned medium used. Standard characterization confirmed the preservation of hESC properties, but the feeder-free cultures could not be maintained longer than 37 passages. The early differentiation process was characterized in the short term after switching hESCs cultured on feeders to feeder-free conditions. Transmission electron microscopy showed an epithelium-like structure inside the hESC colonies, whereas the peripheral cells revealed the acquisition of a rather mesenchymal-like phenotype. Immunochemistry analysis showed that cells at the periphery of the colonies had a negative E-cadherin expression and a positive Vimentin expression, suggesting an epithelial–mesenchymal transition (EMT). Nuclear staining of ß-catenin, positive N-cadherin and negative Connexin 43 expression were also found in the mesenchymal-like cell population. After RT–PCR analysis, Slug and Snail, both EMT-related transcription factors, were detected as up-regulated in the mesenchymal-like cell population. Taken together, our data suggest that culturing hESCs in feeder-free conditions enhances an early differentiation process identified as an EMT.

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