Mol. Hum. Reprod. Advance Access published online on November 7, 2006
Molecular Human Reproduction, doi:10.1093/molehr/gal091
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1 Research Centre Reproduction and Genetics, University Hospital and Medical School of the Vrije Universiteit Brussel (VUB, Free University of Brussels), Brussels, Belgium
* To whom correspondence should be addressed. 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
Received September 1, 2006
Revised September 29, 2006
Accepted October 6, 2006
Article
Epithelial-mesenchymal transition process in human embryonic stem cells cultured in feeder-free conditions
U. Ullmann 1 *, P. In’t Veld 2, C. Gilles 3, K. Sermon 4, M. De Rycke 4, H. Van de Velde 5, A. Van Steirteghem 5, and I. Liebaers 4
2 Experimental Pathology Department, Medical School of the VUB, Brussels, Belgium
3 Laboratory of Developmental and Tumour Biology, University of Liège, Brussels, Belgium
4 Research Centre Reproduction and Genetics, University Hospital and Medical School of the Vrije Universiteit Brussel (VUB, Free University of Brussels) , Brussels, Belgium; Centre for Medical Genetics, University Hospital of the VUB, Brussels, Belgium
5 Research Centre Reproduction and Genetics, University Hospital and Medical School of the Vrije Universiteit Brussel (VUB, Free University of Brussels) , Brussels, Belgium; Centre for Reproductive Medicine, University Hospital of the VUB, Brussels, Belgium
U. Ullmann, E-mail: urielle.ullmann{at}az.vub.ac.be
Abstract 
-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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