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Hum. Reprod. Advance Access originally published online on February 14, 2006
Human Reproduction 2006 21(6):1349-1358; doi:10.1093/humrep/del017
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© The Author 2006. 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
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Cytotrophoblast stem cell lines derived from human embryonic stem cells and their capacity to mimic invasive implantation events

R. Harun1,4, L. Ruban1, M. Matin1,5, J. Draper1, N.M. Jenkins1, G.C. Liew1, P.W. Andrews1, T.C. Li2, S.M. Laird3 and H.D.M. Moore1,6

1 Centre for Stem Cell Biology, University of Sheffield, 2 Royal Hallamshire Hospital, Central Sheffield University Teaching Hospital Trust and 3 Biomedical Research Centre, Sheffield Hallam University, Sheffield, UK

4 Present address: National Population and Family Development Board, Ministry of Women, Family and Community Development, Bangunan LPPKN, 12B Jalan Raja Laut, Peti Surat 10416, 50712 Kuala Lumpur, Malaysia

5 Present address: Department of Biology, Institute of Biotechnology and Tissue Engineering, Ferdowsi University of Mashhad, Mashhad, Iran

6 To whom correspondence should be addressed at: Centre for Stem Cell Biology, University of Sheffield, Western Bank, Sheffield S10 2TN, UK. E-mail: h.d.moore{at}shef.ac.uk

BACKGROUND: An effective embryonic–maternal interaction is crucial for successful human pregnancy. Failure of this process is a major cause of infertility and can lead to placental dysfunction resulting in recurrent miscarriage, fetal retardation and pre-eclampsia. Research is severely constrained by ethical and practical considerations; therefore, we aimed to generate cytotrophoblast stem (CTBS) cell lines from human embryonic stem cells (HESCs). METHOD: beta-HCG was used as a marker of viable trophoblast cells. In defined culture, embryoid bodies were generated from HESCs and selected for trophoblast enrichment by rounds of cellular aggregation and disaggregation. Distinct CTBS cell lines were isolated and characterized. Spheroid cytotrophoblast bodies were generated and their interaction with luteal-phase endometrial stroma was analysed by real-time image analysis. RESULTS: Three CTBS cell lines were derived, which were maintained in the absence of residual HESCs, fibroblast feeder cells or extracellular matrix. CTBS cells displayed typical cytotrophoblast and syncytiotrophoblast characteristics and exhibited further differentiation to invasive endovascular cell phenotype. One cell line was generated with constitutive expression of enhanced green fluorescent protein (eGFP). Spheroid trophoblast bodies mimicked closely the early invasive stages of implantation when incubated with human endometrial stromal preparations in vitro. CONCLUSION: These human CTBS cell lines are a significant new model for investigating human placentation and may have considerable potential in cell therapy applications.

Key words: cytotrophoblast/embryonic stem cells/implantation


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