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Decreases the Viability of Mouse Blastocysts In Vitro and In Vivo1
a University of Louvain Medical School, Physiology of Human Reproduction Research Unit, B-1200 Brussels, Belgium
b Pig Research Institute, Department of Applied Biology, Miaoli, Taiwan
Mouse blastocysts were exposed for 24 h to various concentrations of recombinant mouse tumor necrosis factor 
(TNF) and observed for their capacity to implant in vitro on a fibronectin-coated substrate or to develop in vivo after their transfer into surrogate females. Compared with findings in control blastocysts, exposure to TNF resulted in a significant reduction in the average number of cells in the inner cell mass (ICM) lineage. This effect was associated with a significant increase in the frequency of cells identified as engaged in apoptosis by means of the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling technique. No difference was found in the incidence of nuclear fragmentation between control and TNF-exposed blastocysts. When TNF-pretreated blastocysts were allowed to implant in vitro, significantly fewer embryos were able to maintain a structured ICM cluster at the center of the trophectoderm outgrowth. Although no difference was found in the average surface area of the outgrowths, implants derived from TNF-treated blastocysts contained significantly fewer nuclei than implants from control embryos. After transfer into recipient mice, TNF-pretreated blastocysts implanted at about the same rate as control embryos, but a significantly higher rate of resorption was found among fetuses after exposure to the cytokine. In addition, the weight of the surviving fetuses was significantly lower than for control fetuses. These data indicate that the impact of TNF on blastocysts is specifically aimed at the ICM lineage and that TNF decreases the ability of embryos to differentiate into fetuses after implantation.
2 Correspondence: R. De Hertogh, OBST 5330 Research Unit, University of Louvain Medical School, 53 Avenue Mounier, B-1200 Brussels, Belgium. FAX: 32 2 7645396; dehertogh{at}obst.ucl.ac.be
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