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In vitro  production (IVP) embryos have a reduced quality and poor cryotolerance in comparison to  in vivo  embryos. This study investigated whether free fatty acid (FFA) conditions, fatty acid free (FAF)- synthetic oviduct fluid (SOF) without or with 25 μM of saturated stearic (C 18:0 ) or unsaturated oleic (C 18:1 ) acid during the first 5 IVP days, relate to quality and cryosurvival of day 8 blastocysts. Apart from the blastocyst scores, both 1) number and size of lipid droplets of fresh blastocysts and 2) total number and apoptotic and necrotic cells, before and after freezing-thawing, were scored by confocal microscopy. Blastocyst rates were significantly lower in the FAF SOF condition in comparison to other groups. Interestingly, blastocysts originating from the C 18:1  group, with a significantly higher lipid content, and blastocysts from the FAF SOF group demonstrated a high cryosurvival rate (70.1 and 67.4%, respectively) comparable with  in vivo  blastocysts (68%), in contrast to the poor cryosurvival of C 18:0  exposed embryos (17.6%). In all freeze-thawed embryos the average amount of apoptotic and necrotic cells increased albeit that the C 18:0  condition rates were higher (43.2%) when compared to C 18:1  (26.0%) and FAF SOF conditions (26.5%). The current data show that FFA administered during early embryonic development significantly affect the cryotolerance of blastocysts.

Fatty Acid Supplementation During in vitro Embryo Production Determines Cryosurvival Characteristics of Bovine Blastocysts