Vitrification of Bovine Blastocysts Developed in Royal Jelly-Supplemented Media

Authors : Ali Galip Önal

Pages : 132-139

View : 23 | Download : 7

Publication Date : 2025-06-30

Article Type : Research Paper

Abstract :Vitrification of mammalian embryos is an important process for the long-term preservation of gametes, providing the storage for extended periods. Selection of an optimal culture medium prior to vitrification is also critical to improve post-thaw survival rates. The present study aimed to evaluate the effects of royal jelly (RJ) and fetal bovine serum (FBS) supplementation on bovine embryo development and cryotolerance by evaluating their impact on post-thawing viability, blastocyst survival rates, and cellular integrity following vitrification. Bovine oocytes were matured in vitro in media supplemented with either 10% FBS or 0.625% RJ. Matured oocytes in vitro fertilized (IVF) and presumptive zygotes were further cultured in synthetic oviductal fluid (SOF) supplemented with their respective treatment (FBS or RJ) until day 8. Blastocysts were vitrified using the open-pulled straw (OPS) method with a cryoprotectant solution containing ethylene glycol (EG) and dimethyl sulfoxide (DMSO). Post-thaw survival rates were assessed at 12 and 24 hours. In order to evaluate embryo quality, blastocysts diameter was recorded and total blastocyst cell numbers were determined using Hoechst 33258 staining under a fluorescence microscope. Data were analysed using one-way ANOVA, with Tukey’s HSD test applied for pairwise comparisons. Blastocyst development rates were significantly lower in the RJ-supplemented group (16.3% ± 3.36) compared to the FBS group (24.8% ± 2.58, P 0.05). Although, survival rates remained numerically higher in the RJ group (50.7% ± 3.41) compared to the FBS group (43.5% ± 4.25) at 24 hours, the difference was not statistically significant. Additionally, blastocyst cell counts and diameters were significantly lower in the RJ group than in the FBS group (P < 0.05) at the post-thaw vitrification, suggesting enhanced structural integrity and resilience remains similar to FBS group against cryoinjury. RJ supplementation resulted in a reduced blastocyst development and appeared to have no negative effects on the cryotolerance of embryos, as indicated by similar post-thaw survival rates and improved cellular integrity. The potential protective effects of RJ may be attributed to its bioactive compounds, including antioxidants and collagen-like proteins, which may mitigate oxidative stress and membrane damage during vitrification. These findings suggest that RJ could serve as a promising alternative supplement for improving embryo quality and cryosurvival. However, further research is required to optimize RJ concentration and elucidate its specific protective mechanisms in vitrified embryos.
Keywords : Sığır Embriyosu, Arı sütü, Yavru Sığır Serumu, Vitrifikasyon

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