Development of rabbit preimplantation blastocysts cultured with precultured endometrial tissue

Abstract

Endometrial fragments were explanted from pseudopregnant rabbits 4.5 days after injecting with human chorionic gonadotrophin and were precultured for 2 days in suspension culture in the presence of oestradiol and progesterone equivalent to concentrations in rabbit serum at that stage. Preimplantation blastocysts were obtained at day 6.5 of pregnancy and cultured in the presence or absence of precultured endometrial fragments. Attachment of the trophoblast to the endometrium was prevented by continuous agitation. After 2 and 3 days, specimens were monitored for development in vitro using light and scanning electron microscopy. Although the development of blastocysts was slower in vitro than in vivo in both groups, development was clearly superior in the presence of precultured synchronous endometrial fragments. In the absence of endometrium, the embryonic anlage appeared disordered, particularly in the caudal region, but in the presence of uterine tissue the blastocysts developed much better. Up to nine somites were differentiated; the neural tube had started to close and the various parts of the brain anlage showed incipient differentiation. Syncytiotrophoblast differentiated in the presence or absence of endometrium in the embryonic and abembryonic hemispheres, but typical patterns were maintained better and cell degeneration was less frequent during co-culture. Although the culture model described here has not been optimized using criteria of blastocyst differentiation, the results suggest that culture of blastocysts with precultured synchronous endometrial fragments is advantageous.