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Human Reproduction, Vol. 18, No. 2, 283-290, February 2003
© 2003 European Society of Human Reproduction and Embryology

An in-vitro model for stromal invasion during implantation of the human blastocyst

Janet Carver, Karen Martin1, Isabella Spyropoulou, David Barlow, Ian Sargent and Helen Mardon2

Nuffield Department of Obstetrics and Gynaecology, University of Oxford, Women’s Centre, John Radcliffe Hospital, Oxford, UK

1 Present address: Section of Reproductive and Developmental Medicine, University of Sheffield, Level 4, Jessop Wing, Sheffield S10 2SF, UK

2 To whom correspondence should be addressed at: Nuffield Department of Obstetrics and Gynaecology, University of Oxford, Women’s Centre, Level 3, John Radcliffe Hospital, Headington, Oxford OX3 9DU, UK. e-mail: hmardon{at}molbiol.ox.ac.uk

BACKGROUND: Implantation failure is likely to be a major cause of infertility. Studies in mice have identified a number of molecules that are involved in implantation, but the mechanisms of implantation in the human remain unclear, largely due to the lack of models for implantation in the human that provide functional information. METHODS: Human hatched blastocysts were co-cultured with human endometrial stromal cell monolayers. Time-lapse photography of implanting blastocysts, immunostaining for cytokeratin and actin, and measurement of hCG secreted into the culture supernatants were performed. RESULTS: Blastocysts attached to and implanted into the stromal cell layer. Trophoblast outgrowth onto, and invasion into, the stromal cell layer occurred largely at two opposite poles, the orientation of which was aligned to that of the stromal cell fibroblasts. High-resolution image analysis demonstrated that the trophoblast completely penetrated the stromal cell layer. Immunostaining of whole-mounts of implantation sites revealed distinctive actin and cytokeratin-positive anchoring structures adjacent to the basal surface of the trophoblast. Blastocysts implanting into stromal cells secreted higher levels of hCG compared with those cultured on plastic. CONCLUSIONS: A robust model for the study of mechanisms of implantation of the human embryo into the endometrial stroma has been established.

Key words: blastocyst/co-culture/endometrial stroma cells/implantation/trophoblast


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