KAMBER DEMİR, Ayşe CAN, Ezgi ERTÜRK, Selin ÖZDEMİRCİ, Hazar KARAÇAM, ASİYE İZEM SANDAL, Gül ÖZTÜRK BAKIRER, RAMAZAN ARICI, Elif Merve ÇINAR, Sema BİRLER, SERHAT PABUCCUOĞLU, KEMAL AK, MİTHAT EVECEN

Effect of different activation techniques on immature and in vitro matured cat oocytes

Bu çalışma olgun olmayan ve in vitro olgunlaştırılmış kedi oositlerinin partenogenetik aktivasyonu üzerine 6-Dimetilaminopurin (6- DMAP) ve Sikloheksimidin (CHX), elektik uyarımı ve kalsiyum ionoforla birlikte farklı kullanım kombinasyonlarının denendiği bir çalışma olarak tasarlandı. İn vitro olgunlaştırma (IVM) aşamasının 44. saatinde polar cisimciği attığı gözlenen oositler olgun (MII), atmayanlar ise olgun olmayan (MI) olarak kabul edildi. Aktivasyon sonrası kültüre aktarılan oositler 48 saat sonra değerlendirildi ve yarıklanmayanlar kültürden çıkarıldı. Yarıklanan oositler/embriyolar Modifiye Sentetik Ovidukt Medyumu (mSOF) içerisinde dört gün daha kültüre devam ettirildi. Kültürün altıncı gününde de embriyolar kaliteleri yönünden değerlendirilerek kaydedildi. Bu çalışma sonuçları göstermiştir ki, (I) gerek olgun, gererekse de olgun olmayan kedi oositleri parthenogenetik aktivasyon sonrasında morula ve blastosist aşamasına ulaşabilmektedir, (II) elektrik ve 6-DMAP’ın birlikte kullanıldığı aktivasyon tekniği, olgun ve olgun olmayan oosit aktivasyonu gruplarının her ikisinde de en başarılı sonuçları vermiştir, (III) çalışmada olgun olmayan oositlerin aktivasyonundan elde edilen morula ve blastosist aşamasındaki embriyolar bu alanda ilktir.

Farklı aktivasyon tekniklerinin olgun olmayan ve in vitro olgunlaştırılmış kedi oositleri üzerine etkisi

This study was conducted to determine the most successful techniques on inmature and in vitro-matured cat oocytes that were parhtenogenically activated using 6-dimethylaminopurine (6-DMAP) and cycloheximide (CHX), in combination with electrical stimulation and calcium ionophore. After 44 h of in vitro maturation, the oocytes with a polar body were separated as mature (M II) and those without a polar body were considered as immature. Four different activation treatments and two control groups were used for parthenogenetic activation with both mature and immature cat oocytes. After 48 h of activation, the oocytes were examined and the non-cleaved oocytes removed. The cleaved oocytes/embryos were cultured in vitro in mSOF medium for an additional four days. After six days of in vitro culture (IVC), embryo quality was evaluated. The results in the present study suggested that (I) both in vitro matured and immature cat oocytes have a potential to develop to morula and blastocyst stages after parthenogenetic activation, (II) electrical stimulation + 6-DMAP is a more useful technique for both matured and immature cat oocytes and (III) to our knowledge, this is the first report that describes morula and blastocyst formation from parthenogenetically activated immature cat oocytes.

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