Intrafallopian transfer of gametes and early stage embryos for in vivo culture in cattle.

It may be possible to avoid inadequate in vitro culture conditions by incubating gametes or embryos in the oviducts for a short time. Ideally, an optimized procedure should be devised, combining in vitro and in vivo systems, in order to achieve synchronization in cattle. We transferred gametes as well as embryos in various stages of development and placed them into the oviducts. Embryos were recovered on Day 7 by flushing of oviducts and uterine horns. Blastocyst rates were determined on Day 7 and on Day 8. Experimental designs included transfer of in vitro matured cumulus oocyte complexes into previously inseminated heifers (COCs group), transfer of in vitro matured COCs simultaneously with capacitated spermatozoa (GIFTs group), transfer of four to eight cell stage embryos developed in vitro after IVM/IVF (Cleaved Stages group) and a group of solely in vitro produced embryos (IVP control group). Our results indicate that in vivo culture of IVM/IVF embryos in the homologous bovine oviduct has a positive influence on subsequent pre-implantation development. In addition, we have evidence that in vitro maturation and in vivo fertilization cannot be synchronized.

Effect of morphological properties of transferred embryonic stages on tubal migration Implications for in vivo culture in the bovine oviduct.

In cattle, there is no practical method, which allows tubal transfer of pre-implantation embryos for routine in vivo culture as it has been established in sheep. The aim of our study was to perform tubal transfer by transvaginal endoscopy in synchronized heifers, in order to expose embryos at various embryonic stages to the physiological mechanisms of migration in the non-ligated oviducts. Various embryonic stages were transferred by transvaginal endoscopy into the oviducts of temporary recipients and were recovered on Day 7. The transfer of embryos in hyaluronate containing medium ("Hyaluronan"), zygotes stripped of cumulus ("Denuded Zygotes"), embryos embedded in cumulus ("Zygotes with Cumulus"), matured oocytes with capacitated spermatozoa ("GIFT") or embryos embedded in Na alginate ("Alginate") led to increasing recovery rates (13, 30, 56, 63 and 71%, respectively). However, the developmental rate on Day 7 was adversely affected (16, 11, 8, 16 and 8%), whereas the blastocyst rate on Day 8 showed more balanced results (17, 14, 18, 21 and 11%). Our data demonstrate that the structural properties of transferred embryos affect tubal migration and are crucial for subsequent in vivo culture. Embryos enclosed in cumulus cells or alginate synchronize more successfully with the oviductal transport systems than denuded stages or embryos in hyaluronate containing medium.

In vivo culture of IVM/IVF embryos in bovine oviducts by transvaginal endoscopy.

This study was conducted to establish a new approach for in vivo culture of in vitro produced embryos in the bovine oviduct by transvaginal endoscopy. Embryos were in vitro matured, fertilized and cultured for 1-4 days and assigned to groups consisting of 10-30 embryos. Embryos were transferred unilaterally into oviducts of 24 heifers by the means of transvaginal endoscopy. After 3-6 days of in vivo incubation embryos were re-collected. Experiment I aimed to evaluate the capability of embryos to migrate to the uterus. The uterine horns of four animals were flushed first, followed by a combined flushing of both oviducts and uterine horns resulting in collection rates of 31 and 34%, respectively. In experiment II, the transfer of embryos into the oviduct close to ovulation (day 1-2--experiment IIA) or at a more advanced cyclic stage (day 3--experiment IIB) succeeded in the collection of 46 and 34% of the transferred complexes, of which 13 and 37% showed the blastocyst stage. This is the first report of successful recovery of transferable blastocysts by transvaginal endoscopy after tubal in vivo culture in the homologous species of originally in vitro produced embryos.