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Hum. Reprod. Advance Access published online on October 20, 2005
Human Reproduction, doi:10.1093/humrep/dei328
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© The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received June 20, 2005
Revised August 29, 2005
Accepted September 5, 2005

Article

Transcriptional profiling of the developmentally important signalling pathways in human embryonic stem cells

Jeung-Yon Rho 1, Kweon Yu 1, Jee-Soo Han 2, Jung-Il Chae 2, Deog-Bon Koo 2, Hyun-Soo Yoon 3, Shin-Yong Moon 4, Kyung-Kwang Lee 2, and Yong-Mahn Han 2*

1 Laboratory of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 52 Eoeun-dong Yuseong-gu, Daejeon 305-806; These authors contributed equally to this work
2 Laboratory of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 52 Eoeun-dong Yuseong-gu, Daejeon 305-806
3 Department of Anatomy and Cell Biology, College of Medicine, Hanyang University, Seoul , Korea
4 Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, Seoul, Korea

* To whom correspondence should be addressed.
Yong-Mahn Han, E-mail: ymhan{at}kribb.re.kr


  Abstract

BACKGROUND: Embryonic stem cells (ESC) maintain their ‘stemness’ by self-renewal. However, the molecular mechanisms underlying self-renewal of human embryonic stem cells (hESC) remain to be elucidated. In this study, expression profiles of the molecules of developmentally important signalling pathways were investigated to better understand the relationships of the signalling pathways for self-renewal in hESC. METHODS: Two human ESC lines were cultured on mouse embryonic fibroblast (MEF) feeder cells. Gene expression was analysed by RT-PCR, realtime RT-PCR and Western blotting. RESULTS: In the bone morphogenetic protein (BMP4), transforming growth factor (TGF-{beta}) and fibroblast growth factor (FGF4) signalling pathways, ligands and antagonists were highly expressed in hESC compared with human embryoid body (hEB). Human ESC showed abundant transcripts of intracellular molecules in the Wnt, Hh and Notch signalling pathways. No difference was detected in the expression level of the JAK/STAT signalling molecules between hESC and hEB. Western blot analysis showed that the transcriptional levels of the signalling molecules in hESC were consistent with translational levels. From the real-time PCR analysis, expression levels of some genes, such as Oct3/4, Nodal and {beta}-catenin, were different between two hESC lines. CONCLUSION: The self-renewal of hESC is probably maintained by coordinated regulation of signallingspecific molecules and in a signalling-specific manner.

Keywords: human embryoid body/human embryonic stem cells/self-renewal/signalling pathways/transcription level.
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