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Lookup NU author(s): Professor Lyle Armstrong, Dr Owen Hughes, Sun Yung, Dr Louise Hyslop, Dr Rebecca Stewart, Ilka Wappler, Dr Heiko Peters, Theresia Walter, Petra Stojkovic, Professor Miodrag Stojkovic, Professor Majlinda LakoORCiD
Understanding the molecular mechanism by which pluripotency is maintained in human embryonic stem cells (hESC) is important for the development of improved methods to derive, culture and differentiate these into cells of potential therapeutic use. Large-scale transcriptional comparison of the hES-NCL1 line derived from a day 8 embryo with H1 line derived from a day 5 embryo (WiCell Inc.) showed that only 0.52% of the transcripts analysed varied significantly between the two cell lines. This is within the variability range that has been reported when hESC derived from days 5-6 embryos have been compared with each other. This implies that transcriptional differences between the cell lines are likely to reflect their genetic profile rather than the embryonic stage from which they were derived. Bioinformatic analysis of expression changes observed when these cells were induced to differentiate as embryoid bodies suggested that quite a few of the downregulated genes were components of signal transduction networks. Subsequent analysis using western blotting, flow cytometry and antibody arrays implicated components of the PI3K/AKT kinase, MAPK/ERK and NFκβ pathways and confirmed that these components are decreased upon differentiation. Disruption of these pathways in isolation using specific inhibitors resulted in loss of pluripotency and/ or loss of viability suggesting the importance of such signalling pathways in embryonic stem cell maintenance. © 2006 Oxford University Press.
Author(s): Armstrong L, Hughes O, Yung S, Hyslop L, Stewart R, Wappler I, Peters H, Walter T, Stojkovic P, Evans J, Stojkovic M, Lako M
Publication type: Article
Publication status: Published
Journal: Human Molecular Genetics
Year: 2006
Volume: 15
Issue: 11
Pages: 1894-1913
ISSN (print): 0964-6906
ISSN (electronic): 1460-2083
Publisher: Oxford University Press
URL: http://dx.doi.org/10.1093/hmg/ddl112
DOI: 10.1093/hmg/ddl112
PubMed id: 16644866
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