Article (Scientific journals)
GSK-3-specific inhibitor-supplemented hESC medium prevents the epithelial-mesenchymal transition process and the up-regulation of matrix metalloproteinases in hESCs cultured in feeder-free conditions
Ullmann, U.; Gilles, Christine; De Rycke, M. et al.
2008In Molecular Human Reproduction, 14 (3), p. 169-79
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Keywords :
epithelial–mesenchymal transition; GSK-3 inhibitor; human embryonic stem cells; feeder-free culture; matrix metalloproteinases
Abstract :
[en] Feeder-free culture induces spontaneous differentiation of human embryonic stem cells (hESCs), identified as an epithelial to mesenchymal transition (EMT). The maintenance of pluripotency of hESCs in feeder-free cultures through the activation of the WNT pathway using a glycogen synthase kinase (GSK)-3-specific inhibitor (BIO) was reported. The aim of this study was to determine the effect of BIO on the EMT process. In contrast with those grown in feeder-free conditions with control medium, hESC colonies cultured with BIO-supplemented hESC medium did not show any fibroblast-like cells at the periphery. Transmission electron microscopy, relative quantitative real-time RT–PCR and immunostaining analyses showed the presence of epithelial features and a diminution of mesenchymal features in the BIO-treated hESCs such as a strong E-cadherin expression, the down-regulation of Vimentin, Snail and Slug expressions and a cytoplasmic β-catenin expression. An up-regulation of matrix metalloproteinases (MMP) MMP-2, MMP-9, MT-1MMP (membrane-type 1 MMP) and EMMPRIN (extracellular MMP inducer) expression was also found associated with the EMT occurring in feeder-free hESCs cultures using mouse embryonic fibroblasts conditioned medium (MEF CM). The presence of BIO clearly down-regulated the expression of these MMPs. This study showed that BIO, a GSK-3-specific inhibitor, prevents the EMT process which is associated with the feeder-free hESC culture. Nevertheless, BIO was not sufficient to expand hESCs in a long-term culture system.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Ullmann, U.
Gilles, Christine ;  Université de Liège - ULiège > Département des sciences cliniques > Labo de biologie des tumeurs et du développement
De Rycke, M.
Van de Velde, H.
Sermon, K.
Liebaers, I.
Language :
English
Title :
GSK-3-specific inhibitor-supplemented hESC medium prevents the epithelial-mesenchymal transition process and the up-regulation of matrix metalloproteinases in hESCs cultured in feeder-free conditions
Publication date :
2008
Journal title :
Molecular Human Reproduction
ISSN :
1360-9947
eISSN :
1460-2407
Publisher :
Oxford University Press, Oxford, United Kingdom
Volume :
14
Issue :
3
Pages :
169-79
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 22 June 2010

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