1 AUTOCRINE COMMUNICATION BETWEEN BOVINE EMBRYOS CULTURED IN Primo Vision DISHES® OUTWEIGHS POSSIBLE NEGATIVE INFLUENCES OF BAD EMBRYOS

Abstract

Group culture is being used extensively for mammalian embryos, but has not been adopted so far in human embryo culture. Some doubts about its possible benefits remain because it has been hypothesised that bad quality embryos might have a negative effect on other embryos. New group culture devices have been designed allowing individual follow-up of embryos, such as Primo Vision dishes^® (well of-the-well for 10 embryos in group culture; Cryo-Innovation, Budapest, Hungary). By using Primo Vision dishes^®, we investigated the influence of the developmental stage of the neighbours and co-cultured embryos on the outcome at 192 h post-insemination (hpi) of a particular embryo compared with its individually cultured counterparts. Bovine presumed zygotes (n = 789; 4 replicates) produced in vitro were randomly allocated to Primo Vision dishes^® or individual culture in SOF supplemented with 0.4% BSA and insulin, transferrin, selenium (Wydooghe et al. 2013 Reprod. Fertil. Dev. Epub). Cleavage rate was checked at 45 hpi: 5- to 8-cell embryos were classified as fast embryos and 2- to 4-cell embryos as slow embryos. Blastocyst development was evaluated at 192 hpi. Moreover, to evaluate which embryos benefited most from being in group (fast or slow embryos), we looked retrospectively at the influence of the developmental stage of the neighbours and the co-cultured embryos on blastocyst development. This was done separately for slow and fast embryos compared with their individually cultured counterparts. Statistical analysis was done using a binary logistic regression model, with group and replicate as fixed effects. Blastocyst development in Primo Vision dishes^® was significantly better than individual culture (39.0% v. 28.5%). This beneficial outcome was mainly caused by a higher blastocyst development of slow embryos. A markedly higher percentage of slow embryos developed into a blastocyst at 192 hpi if they were surrounded by many embryos that also developed into a blastocyst, compared with individually cultured slow embryos (odds ratio: 3.0). In this study, we showed that embryos that were not cleaved at 45 hpi did not negatively affect the potential of their neighbours to become a blastocyst at 192 hpi, regardless of whether the embryo in question was a fast or a slow embryo. However, when fast embryos were in a less than favourable environment, meaning that less than 30% of their co-cultured embryos reached the blastocyst stage, blastocyst development was compromised compared with individual culture of fast embryos (odds ratio: 0.3). From our results, we clearly show that Primo Vision dishes^® can combine the benefits of group culture (autocrine communication) and individual culture (individual follow-up). Taking fast embryos out of the Primo Vision dish^® for further individual culture while slow embryos remain in group is a possible approach to increase total blastocyst rates.