Journal Article

Epithelial–mesenchymal transition process in human embryonic stem cells cultured in feeder-free conditions

U. Ullmann, P. In’t Veld, C. Gilles, K. Sermon, M. De Rycke, H. Van de Velde, A. Van Steirteghem and I. Liebaers

in MHR: Basic science of reproductive medicine

Published on behalf of European Society of Human Reproduction and Embryology

Volume 13, issue 1, pages 21-32
Published in print January 2007 | ISSN: 1360-9947
Published online November 2006 | e-ISSN: 1460-2407 | DOI: http://dx.doi.org/10.1093/molehr/gal091
Epithelial–mesenchymal transition process in human embryonic stem cells cultured in feeder-free conditions

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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 β-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.

Keywords: human embryonic stem cells; feeder-free culture; matrigel; differentiation; epithelial–mesenchymal transition

Journal Article.  8155 words.  Illustrated.

Subjects: Reproductive Medicine

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