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Hum. Reprod. Advance Access published online on September 30, 2004
Human Reproduction, doi:10.1093/humrep/deh497
© 2004 by European Society of Human Reproduction and Embryology
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Received January 5, 2004
Revised April 13, 2004
Accepted August 4, 2004

Article

Embryotrophic factor-3 from human oviductal cells enhances proliferation, suppresses apoptosis and stimulates the expression of the {beta}1 subunit of sodium-potassium ATPase in mouse embryos

J.S. Xu 1, Y.L. Lee 1, K.F. Lee 1, K.L. Kwok 1, W.M. Lee 2, J.M. Luk 3, and W.S.B. Yeung 1*

1 Department of Obstetrics and Gynaecology, Queen Mary Hospital, The University of Hong Kong, Hong Kong, China
2 Department of Zoology, Queen Mary Hospital, The University of Hong Kong, Hong Kong, China and
3 Department of Surgery, The University of Hong Kong, Pokfulam Road, Hong Kong SAR, China

* To whom correspondence should be addressed. E-mail: wsbyeung{at}hkucc.hku.hk.


  Abstract

BACKGROUND: Embrytrophic factor-3 (ETF-3) from human oviductal cells enhanced the development of mouse preimplantation embryos. This report studied the embryotrophic mechanisms of the molecule. METHODS AND RESULTS: Mouse embryos were incubated with ETF-3 for 24 h at different stages of development. ETF-3 treatment between 96 and 120 h post-HCG increased the cell count of blastocysts, whilst treatment between 72 and 96 h post-HCG enhanced the expansion and hatching of the blastocysts. ETF-3 increased the cell number of the embryos by suppressing apoptosis and increasing proliferation as determined by TUNEL and bromodeoxyuridine uptake assays, respectively. Real-time quantitative PCR showed that the in vivo developed and ETF-3-treated blastocysts had a significantly higher mRNA copy number of Na/K-ATPase-{beta}1, but not of hepsin, than that of blastocysts cultured in medium alone. The former gene was associated with cavitation of blastocysts while the latter was related to hatching of blastocyst. The beneficial effect of ETF-3 on blastocyst hatching was also seen when ETF-3-supplemented commercially available sequential culture medium for human embryo culture was used to culture mouse embryos. CONCLUSIONS: ETF-3 improves embryo development by enhancing proliferation, suppressing apoptosis and stimulating expression of genes related to blastocyst cavitation. Supplementating human embryo culture medium with ETF-3 may improve the success rate in clinical assisted reproduction.

Keywords: apoptosis; embryotrophic factor; oviduct; proliferation; sodium-potassium ATPase.
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