Mustafa Numan BUCAK, Muharrem SATILMIŞ, SEDAT HAMDİ KIZIL, TAHİR KARAŞAHİN, NUMAN AKYOL

Sığır embriyolarının in vitro gelişiminde kültür medyumlarına katılan antioksidanların etkisi

Bu çalışmanın konusu, in vitro sığır embriyolarının gelişiminde kültür medyumlarına katılan antioksidanların etkisinin araştırılması olmuştur. Lokal mezbahadan sağlanan inek ovaryumları fizyolojik tuzlu su (%0.9), antibiyotik içeren taşıma sıvısı içerisinde ve 30°C’lik sabit sıcaklık sağlayan termosta 2-3 saat içerisinde laboratuvara getirilmiştir. Oositlerin in vitro maturasyonu, fertilizasyonu sonrası, farklı antioksidanların (sisteamin 25 μM, sisteamin 50 μM; dithioerythritol 25 μM, dithioerythritol 50 μM; glutatyon 5 μM, glutatyon 10 μM) ilave edildiği Charles Rosencrans (CR1aa) ve Sentetik Ovidukt Sıvısı (SOF) embriyo kültür medyumlarında kültür periyoduna alınarak embriyo gelişim aşamaları takip edilmiştir. Glutatyon (5 ve 10 μM) içeren CR1aa kültür medyumunda, diğer gruplara nazaran ekspand blastosist aşamasında embriyo elde edilememiş, bu fark ise önemli sayılmıştır (P0.05). Farklı antioksidanlar içeren SOF kültür medyumunda, dithioerythritol (50 μM)’un antioksidan içermeyen gruba göre, bölünmüş embriyo oranında önemli artışlar sağladığı görülmüştür (P

Anahtar Kelimeler:

sığır, embriyonik gelişim, embriyo, antioksidanlar, in vitro kültür, kültür ortamı

Effect of antioxidants added to culture on the in vitro development of bovine embryos

The objective of the present study was to investigate the effect of antioxidants added to culture media on the development of in vitro bovine embryos. Cow ovaries provided from local slaughterhouses were transported to the laboratory in transport medium, consisting of physiological saline (0.9%) and antibiotics, at 30°C within 2-3 hours. Following their in vitro maturation and fertilization, the oocytes were cultured in Charles Rosenkrans medium (CR1aa) and synthetic oviduct fluid (SOF) medium containing different antioxidants (25 μM cysteamine, 50 μM cysteamine; 25 μM dithioerythritol, 50 μM dithioerythritol; 5 μM glutathione, 10 μM glutathione) and were monitored for embryonic development. In the CR1aa culture medium containing glutathione (5 and 10 μM), the expanded blastocyst stage was not obtained, compared to the other groups, and this difference was considered to be significant (P<0.05). The percentages of embryos reaching the stages of morula, blastocyst and hatched blastocyst decreased insignificantly in the groups with glutathione. As regards percentages of cleavage embryos, cleavage and development of blastocyst embryos from cultured oocytes, CR1aa medium with dithioerythritol (50 μM) gave higher results than the other groups, but this difference was statistically insignificant (P>0.05). Upon the investigation of SOF culture medium containing different antioxidants, it was determined that dithioerythritol (50 μM) provided significant increase in the percentage of cleavage embryos (P<0.05), compared to the group with no antioxidant. Embryos did not reach the expanded blastocyst and hatched blastocyst stages in the groups with dithioerythritol (25 μM) and glutathione (5 and 10 μM). Statistically, differences did not exist in the percentages of embryos reaching the morula, and expanded and hatched blastocyst stages among study groups. Furthermore, in the present study, development rates of embryos were compared in culture media of CR1aa and SOF. It was determined that CR1aa medium produced higher percentages of embryonic developmental stages, excluding percentages of embryos reaching the hatched blastocyst stage, in comparison to SOF medium (P<0.01).

Keywords:

cattle, embryonic development, embryos, antioxidants, in vitro culture, culture media,

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