Effects of Cysteamine on Sheep Embryo Cleavage Rates

Abstract

Oxidative stress during in vitro culture leads to defects in development of gametes and embryos. Several antioxidants such as cysteamine, L-ascorbic acid, beta mercaptoethanol, cysteine, glutathione, proteins, vitamins have been used to supplement culture media to counter the oxidative stress. This study was conducted to detect the effect of adding cysteamine to the maturation medium to subsequent cleavage rates of sheep embryos. Totally 604 ovaries were obtained by ten replica and 2060 oocytes were collected. The cumulus oocyte complexes were recovered by the slicing method. A total of 1818 selected oocytes were divided into two groups and used for maturation (88.25%). The first group was created as supplemented with cysteamine (Group A) and second group (Group B, control) without cysteamine in TCM-199. The two groups were incubated for 24 h at 38.8 °C in an atmosphere of 5% CO2 in humidified air for in vitro maturation (IVM). After IVM, oocytes were fertilized with 50 x 107 / mL fresh ram semen in BSOF medium for 18 h. After fertilization, maturation groups were divided into two subgroups with different culture media: Group AI-SOF (Synthetic Oviduct Fluid medium), Group AII-CR1aa (Charles Rosencrans medium), Group BI-SOF and Group BII-CR1aa were achieved. Cleavage rates were evaluated at day 2. post insemination. The rates of cleavage were detected as 59.54% (184/309), 55.44% (173/312), 65.34% (215/329), 59.34% (200/337) respectively, with showing no statistically significant difference between the groups at the level of P>0.05. In conclusion, supplementing cysteamine to maturation media in TCM-199 did not affect the cleavage rates of sheep embryos in SOF and CR1aa culture media.

Keywords

• Sheep, cysteamine, maturation, cleavage rate, embryo

Sisteaminin Koyun Embriyo Bölünme Oranlarına Etkileri

Abstract

In vitro kültür esnasında oksidatif stres gametlerin ve embriyoların gelişiminde hasarlara yol açar. Sisteamin, L-askorbik asit, beta merkaptoetanol, sistein, glutatyon gibi birçok antioksidanlar, proteinler, vitaminler oksidatif stresi önlemek için kültür medyumlarına eklenmiştir. Bu çalışma olgunlaşma medyumuna sisteamin ilavesinin koyun embriyonik bölünme oranları üzerine etkisinin saptanması için tasarlandı. Toplam 604 ovaryum 10 kerede toplandı ve 2060 koyun oositi elde edildi, doğrama metodu ile 1818 adet oosit kumulus kompleksleri seçilerek olgunlaşma amacıyla iki gruba ayrıldı (%88,25). İlk grup (Grup A) TCM-199 medyumuna sisteamin eklenen oositler, diğer grup (Grup B, kontrol) TCM-199 medyumuna sisteamin eklenmeyen oositler olarak oluşturuldu. İki grupta 24 saat boyunca 38,8 °C’de %5 CO2 altında, nemli havada in vitro maturasyon (IVM) için inkübe edildi. Oositler, IVM sonrası 50 x 107 / mL taze koç sperması ile BSOF medyumunda 18 saat süreyle döllendi. Döllenme sonrası, olgunlaşma grupları değişik kültür medyumları ile iki alt gruba ayrıldı ve Grup AI-SOF (Sentetik Ovidukt Sıvısı Medyumu), Grup AII-CR1aa (Charles Rosencrans Medyumu), Grup BI-SOF ve Grup BII-CR1aa elde edildi. Bölünme oranları döllenme sonrası 2. günde değerlendirildi. Bölünme oranları gruplar arasında istatistik açıdan önemli fark olmadan (P>0,05) sırasıyla, %59,54 (184/309), %55,44 (173/312), %65,34 (215/329), %59,34 (200/337) olarak saptandı. Sonuç olarak, TCM-199 olgunlaşma medyumuna sisteamin eklenmesinin SOF ve CR1aa kültür medyumlarındaki koyun embriyo bölünme oranları üzerine bir etkisinin olmadığı saptandı.

Keywords

• Koyun, sisteamin, olgunlaşma, bölünme oranı, embriyo

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