Oil-free culture system for in vitro bovine embryo production

The use of oil to avoid water evaporation from cell culture has several disadvantages, amongst which there is the migration of compounds from media to oil and from oil to media. The aim of this study was to evaluate the osmolality of a culture system using four-well plates with water in the central hole as an alternative to in vitro bovine embryo production (IVP). In addition, the osmolality changes of the oocyte washing medium were assessed in 35mm dishes with or without 2 mL of silicon oil overlay. Osmolality of oocyte washing medium changed a great deal over time after 60 minutes on a 39&deg;C heated plate (291 mOsm kg-1), which was not detected when the medium was overlaid with silicon oil (280 mOsm kg-1; P<0.05). During the maturation period, the presence of water in the central hole of four-well plates maintained the osmolality in the same pattern as oil overlay (293&plusmn;0 vs 294&plusmn;1.8 mOsm kg-1; P>0.05). Blastocyst rates were higher when embryos were cultured in presence of water or oil (29.7 and 29.9% for water and 33% in oil conventional microdrop system), except in the group that oocytes were washed in hyperosmotic washing medium (15.1%; P<0.05). Groups cultured in absence of water in the central hole had lower blastocyst rates (P<0.05) independently of exposure (15.5%) or not (16.2 and 16.8%) to hyperosmotic washing medium. In conclusion, four-well plates with water in the central hole can be an alternative to replace oil overlay for bovine IVP, maintaining stable osmolality and embryo development rates.

Abstract licence: CC BY-NC

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