Vitrification of Bovine Blastocysts Developed in Royal Jelly-Supplemented Media

Ali Galip Önal

Makale detayları

Vitrification of Bovine Blastocysts Developed in Royal Jelly-Supplemented Media

Yazarlar:

Ali Galip Önal

Yayıncı:

Cukurova University

Ana Başlıklar:

Zootekni | Genetik ve Biyoistatistik

Dergi:

Çukurova Tarım ve Gıda Bilimleri Dergisi Vol.40 Issue 1, pp.132-139

Belge Tipi:

Makale

Yayın Tarihi:

6/2025

Belge Dili:

İngilizce

Document ID (Org No.):

2001353364

pISSN:

2636-7874

Anahtar kelimeler:

Özet:

 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 Tukeys 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, suggesting that RJ may affect early embryonic development. Despite the decreased blastocyst development, RJ-derived blastocysts exhibited comparable post-thaw survival rates to their FBS counterparts at 12 hours FBS-V: 51.3  3.72 vs. RJ-V: 48.3  4.09, 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.

 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 Tukeys 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, suggesting that RJ may affect early embryonic development. Despite the decreased blastocyst development, RJ-derived blastocysts exhibited comparable post-thaw survival rates to their FBS counterparts at 12 hours FBS-V: 51.3  3.72 vs. RJ-V: 48.3  4.09, 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.

 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 Tukeys 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, suggesting that RJ may affect early embryonic development. Despite the decreased blastocyst development, RJ-derived blastocysts exhibited comparable post-thaw survival rates to their FBS counterparts at 12 hours FBS-V: 51.3  3.72 vs. RJ-V: 48.3  4.09, 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.

 Memeli embriyolarının vitrifikasyonu, gametlerin uzun dönem korunması için önemli bir tekniktir. Vitrifikasyon öncesinde en uygun kültür ortamının seçimi, embriyoların çözme sonrası hayatta kalma oranlarını artırmak için hayati önem taşımaktadır. Bu çalışma, Arı sütü AS ve fetal sığır serumu FBS takviyesinin sığır embriyo gelişimi ve kriyotolerans üzerindeki etkilerini inceleyerek, çözme sonrası canlılık, blastosistlerin hayatta kalma oranları ve hücresel bütünlükleri üzerindeki etkilerini araştırmayı amaçlamaktadır. Sığır oositleri, kültür ortamında 10 FBS veya 0.625 AS takviyesi ile in vitro olgunlaştırılmıştır. In vitro fertilizasyon IVF işleminin ardından oluşan zigotlar 8. güne kadar FBS veya AS ila edilen sentetik oviduktal sıvı SOF içinde kültüre edilmiştir. 8. Güne gelen Blastosistler, açık uçlu çekilmiş pipetler OPS yöntemi ile etilen glikol EG ve dimetil sülfoksit DMSO içeren bir kriyoprotektan solüsyon kullanılarak vitrifike edilmiştir. Çözme sonrası hayatta kalma oranları 12 ve 24 saat sonra değerlendirilmiştir. Embriyo kalitesi, blastosist çapı ve toplam hücre sayısı temelinde Hoechst 33258 boyama yöntemi kullanılarak floresan mikroskop altında belirlenmiştir. Elde edilen ANOVA ile analiz edilmiş ve gruplar Tukey HSD testi ile karşılaştırılmıştır. Blastosist gelişim oranları, AS ile desteklenen grupta 16.3  3.36 FBS grubuna kıyasla 24.8  2.58, P  0.05 anlamlı olarak daha düşük bulunmuştur. Bu durum, ASnin erken embriyonik gelişimi etkileyebileceğini göstermektedir. Bununla birlikte, azalan blastosist gelişimine rağmen, AS ile kültür edilen blastosistlerin çözme sonrası 12 saatlik hayatta kalma oranları, FBS grubuyla benzer seviyede bulunmuştur FBS-V: 51.3  3.72 vs. RJ-V: 48.3  4.09, P  0.05. 24 saatlik hayatta kalma oranları ise AS grubunda sayısal olarak daha yüksek olmasına rağmen RJ: 50.7  3.41, FBS: 43.5  4.25, bu fark istatistiksel olarak anlamlı bulunmamıştır. Ayrıca, blastosist hücre sayıları ve çapları AS grubunda FBS grubuna kıyasla anlamlı olarak daha düşük bulunmuştur P  0.05. Elde edilen sonuçlar, ASnin embriyolarda yapısal bütünlüğü olumsuz etkilemediğini ve kriyozarara karşı FBS grubu ile benzer etki gösterdiğini düşündürmektedir. AS takviyesi, blastosist gelişim oranını düşürmesine rağmen kriyotoleransı olumsuz etkilememiştir. Çözme sonrası hayatta kalma oranlarının korunması ve hücresel bütünlüğün benzer kalması, ASnin embriyolar üzerindeki olası koruyucu etkilerini işaret etmektedir. Bu koruyucu etkiler, ASnin içerdiği antioksidanlar ve kollajen benzeri proteinler gibi biyoaktif bileşiklere bağlanabilir ve vitrifikasyon sırasında oluşabilecek oksidatif stres ile membran hasarını azaltabilir. Elde edilen bulgular, ASnin embriyo kalitesini ve donma sonrası hayatta kalma oranları artırmada potansiyel bir alternatif takviye olarak değerlendirilebileceğini göstermektedir. Bununla birlikte, ASnin optimum konsantrasyonunun belirlenmesi ve vitrifikasyon sürecinde spesifik koruyucu mekanizmalarının aydınlatılması için daha fazla araştırmaya ihtiyaç duyulmaktadır.

Makale detayları

Vitrification of Bovine Blastocysts Developed in Royal Jelly-Supplemented Media

Yazarlar:

Ali Galip Önal

Yayıncı:

Cukurova University

Ana Başlıklar:

Zootekni | Genetik ve Biyoistatistik

Dergi:

Çukurova Tarım ve Gıda Bilimleri Dergisi Vol.40 Issue 1, pp.132-139

Belge Tipi:

Makale

Yayın Tarihi:

6/2025

Belge Dili:

İngilizce

Document ID (Org No.):

2001353364

pISSN:

2636-7874

Bovine Embriyo

Royal Jelly

Fetal Bovine Serum

VitrificationSığır Embriyosu

Arı sütü

Yavru Sığır Serumu

Vitrifikasyon

Sayı

Makale detayları

Yazarlar: Ali Galip Önal

Yayıncı: Cukurova University

Ana Başlıklar: Zootekni | Genetik ve Biyoistatistik

Dergi: Çukurova Tarım ve Gıda Bilimleri Dergisi Vol.40 Issue 1, pp.132-139

Belge Tipi: Makale

Yayın Tarihi: 6/2025

Belge Dili: İngilizce

Document ID (Org No.): 2001353364

pISSN: 2636-7874

Sayı

Yazarlar:

Ali Galip Önal

Yayıncı:

Cukurova University

Ana Başlıklar:

Zootekni | Genetik ve Biyoistatistik

Dergi:

Çukurova Tarım ve Gıda Bilimleri Dergisi Vol.40 Issue 1, pp.132-139

Belge Tipi:

Makale

Yayın Tarihi:

6/2025

Belge Dili:

İngilizce

Document ID (Org No.):

2001353364

pISSN:

2636-7874