The in vitro development of zygotes of domestic species to the blastocyst stage is facilitated by culture in groups, suggesting a role for autocrine/paracrine factors. A novel method was used to investigate the potential role of such factors using in-vitro-produced and in-vivo-derived porcine embryos. The development of individual zygotes to the blastocyst stage was optimal when they were cultured 81-160 mum apart. As the distance between the embryos was increased, blastocyst rates declined significantly, reaching zero beyond 640 mum. Blastocyst volume and cell number (both inner cell mass and trophectoderm) were also increased when the distance apart was between 81 and 160 mum. Culturing embryos in groups at different stages of development suggested that group culture confers a greater advantage to development after the activation of the genome. Group culture of in-vivo-derived embryos showed a weak distance effect. The results suggest a role for as yet unknown diffusible paracrine/autocrine factors released by early porcine embryos in promoting the growth of neighbouring embryos in vitro. This advantage is observed to a lesser extent by in-vivo-derived zygotes which are likely to have been better conditioned for development in vitro by being conceived in the female reproductive tract.

The data suggests that cryopreservation in IVF is a safe procedure and that amino acid turnover can be used to select which cryopreserved embryo will develop to the blastocyst stage, irrespective of their post-thaw grade.

No abstract