BOEC-Exo Addition Promotes In Vitro Maturation of Bovine Oocyte and Enhances the Developmental Competence of Early Embryos

Abstract

Simple Summary The results of the present study proved that the addition of bovine oviductal epithelial cell derived exosomes (BOEC-Exo) to the in vitro maturation (IVM) media improved the bovine oocyte maturation and early embryo development. The addition of BOEC-Exo not only significantly enhanced the polar body exclusion, but also enhanced the expression of connexins in cumulus oocyte complexes (COCs). Likewise, the reactive oxygen species (ROS) level, protein expressions of SIRT-1, and mitochondrial membrane potential (Delta psi m) also suggested that BOEC-Exo addition to IVM media is highly beneficial for in vitro bovine oocyte maturation. Furthermore, BOEC-Exo treatment to the primary cultured bovine cumulus cells significantly attenuated apoptosis, which also showed its positive influence on the COCs. Moreover, oocytes that were matured in the presence of BOEC-Exo led to the production of a significantly higher quantity and quality of day-8 blastocysts. Additionally, the BOEC-Exo treated blastocysts had a higher implantation potential when compared with the control. Our results suggest that the addition of BOEC-Exo to IVM media significantly enhanced the percentage of oocytes maturation and improved the embryo quantity and quality. Exosomes are nano-sized vesicles with abundant nucleic acids, proteins, lipids, and other regulatory molecules. The aim of this study was to examine the effect of BOEC-Exo on bovine in vitro oocyte maturation and in vitro embryo development. We found that a 3% Exo supplementation to IVM media significantly enhanced the oocyte maturation and reduced the accumulation of ROS in MII-stage bovine oocytes. Oocyte maturation related genes (GDF9 and CPEB1) also confirmed that 3% Exo treatment to oocytes significantly (p < 0.05) enhanced the oocyte maturation. Next, we cultured bovine cumulus cells and assessed the effects of 3% Exo, which showed a reduced level of apoptotic proteins (caspase-3 and p-NF-kappa B protein expressions). Furthermore, we examined the gap junction (CX43 and CX37) and cumulus cells expansion related genes (HAS2, PTX3, and GREM1) in cumulus-oocyte complexes (COCs), and all those genes showed significantly (p < 0.05) higher expressions in 3% Exo-treated COCs as compared with the control group. Moreover, peroxisome proliferator-activated receptors (PPARs) and lipid metabolism-related genes (CPT1 and FABP3) were also analyzed in both the control and 3% Exo groups and the results showed significant (p < 0.05) enhancement in the lipid metabolism. Finally, the oocytes matured in the presence of 3% Exo showed a significantly higher rate of embryo development and better implantation potential. Finally, we concluded that Exo positively influenced bovine oocyte in vitro maturation and improved the early embryo's developmental competence.