Supplementation of bovine follicular fluid during in vitro maturation increases oocyte cumulus expansion, blastocyst developmental kinetics, and blastocyst cell number.

Abstract Bovine follicular fluid (bFF) is the natural milieu for oocyte growth and development. However, its value as supplementation to in vitro maturation medium is still questioned due to inconsistent results. In this study we hypothesized that adding 10% of follicular fluid as well as heat treating it to inhibit the complement system, would produce higher quality embryos. To do so, experiments were conducted to compare the effect of bFF and heat-treated bFF (bFFin) on oocyte competence assessed by different parameters such as nuclear and cytoplasmic maturation, IVF efficiency, in vitro embryo development and embryo survivability post-vitrification. No differences on nuclear maturation nor cortical granules migration were observed but differences were found on oocyte's cumulus cell expansion, with bFF group having the highest increase (79.0 ± 3.7%). bFFin had a negative impact on IVF efficiency (58.6 ± 3.2%), but no differences were found between bFF (62.9 ± 3.2%) and control (72.8 ± 3.0%). Although the cleavage and blastocyst rate were similar between groups, the day 6 embryo development rate was higher in bFFin group, suggesting an accelerated developmental kinetics. Hatched blastocysts from the bFF group showed a higher cell count than the control group (241.3 ± 20.1 and 185.8 ± 10.0, respectively), and bFFin embryos showed values in between (214.9 ± 14.0). No difference on survivability post-vitrification was found between groups, although the blastocyst stage had a significant impact on the survival rate across all groups. In conclusion, using bFF as supplementation to maturation medium showed a higher benefit when comparing to the standard supplementation by having oocytes with higher cumulus expansion rate, faster development of embryos and higher number of cells per embryo. Inactivation of bFF lowered IVF efficiency but didn't compromise blastocyst development and quality. Highlights • The supplementation of maturation medium with bovine follicular fluid shows higher expansion of cumulus cells in oocytes. • The supplementation of maturation medium with bovine follicular fluid shows higher total cell number in in vitro grown embryos. • Inactivation of follicular fluid decreases IVF efficiency but not blastocyst yield. • Inactivation of follicular fluid promotes earlier development of embryos on day 6 of culture. • Bovine follicular fluid is a better alternative to foetal bovine serum supplementation during in vitro maturation of bovine oocytes. [ABSTRACT FROM AUTHOR]