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İn Vitro Olgunlaştırma Medyumunda Fötal Buzağı Serumu Yerine İki Farklı Sentetik Serumun Kullanılması: Sığır Oositlerinin İn Vitro Olgunlaştırma, Fertilizasyon Ve Sonraki Gelişimleri Üzerindeki Etkileri

Replacement of Fetal Calf Serum With Two Different Synthetic Sera in in Vitro Maturation Medium: Effects On Maturation, Fertilization and Subsequent Development of Cattle Oocytes in Vitro

Journal Name:

  • Uludag University Journal of The Faculty of Veterinary Medicine

Publication Year:

  • 2010

Key Words:

Keywords (Original Language):

Author NameUniversity of AuthorFaculty of Author
Abstract (2. Language): 
The aim of this study was to investigate the effect of using synthetic sera (synthetic serum substitute, SSS and serum replacement 1, SR1) instead of fetal calf serum (FCS) in in vitro maturation medium on in vitro maturation (IVM), fertilization (IVF) and subsequent development of bovine oocytes. Selected oocytes collected from ovaries obtained from a local slaughterhouse were matured in tissue culture medium 199 (TCM-199) supplemented with 2 mM glutamine, 0.25 mM Na-pyruvate, 0.5 g/ml FSH, 5 g/ml LH, 100 U/ml penicillin, 100 μg/ml streptomycin and 10% FCS, SSS or SR1 (FCS, SSS and SR1 groups, respectively) for 22 hours. Matured oocytes were in vitro fertilized using frozen bull sperm. Fertilization day was considered as day 0 in the present study. Eighteen hours after IVF, fertilized oocytes were transferred into synthetic oviduct fluid (SOF) culture medium for in vitro culture. Two cell, 8-cell, morulae and blastocyst numbers were recorded. A total of 238, 243 and 200 oocytes were used for FCS, SSS and SR1 groups, respectively. In FCS, SSS and SR1 groups, 179 (75.21%), 177 (72.84%) and 80 (40.00%); 121 (50.84%), 114 (46.91%) and 38 (19.00%); 97 (40.76%), 98 (40.03%) and 21 (10.50%); and 71 (29.83%), 69 (28.40%) and 10 (5.00%) oocytes developed to 2-cell, 8-cell, morulae and blastocyst stage embryos, respectively. The development rates of SR1 group at all recorded stages were significantly lower than those of FCS and SSS groups. There was no significant difference between FCS and SSS groups in terms of the developmental rates at any stage recorded. As a result, SSS is a good alternative for FCS replacement in in vitro maturation of bovine oocytes, however, SR1 is not a good alternative for FCS replacement in in vitro maturation medium.
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Abstract (Original Language): 
Bu çalışmanın amacı sığır oositlerinin in vitro olgunlaştırılması, fertilizasyonu ve sonraki gelişimleri üzerinde olgunlaştırma medyumunda fötal buzağı serumu (FCS) yerine sentetik serumların (synthetic serum substitute, SSS and serum replacement 1, SR1) kullanılmasının etkilerini incelemektir. Yerel bir mezbahadan elde edilen ovaryumlardan kazanılan seçilmiş oositler 2 mM glutamin, 0,25 mM Na-piruvat, 0,5 g/ml FSH, 5 g/ml LH, 100 U/ml penisilin, 100 μg/ml streptomisin ve %10 FCS, SSS ya da SR1 (sırasıyla FCS, SSS ve SR1 grupları) ile takviye edilmiş doku kültür medyumu (TCM-199) içerisinde 22 saat süreyle olgunlaştırılmıştır. Olgunlaşan oositler dondurulmuş boğa spermiyle in vitro fertilize edilmiştir. Fertilizasyon günü bu çalışmada 0. gün olarak kabul edilmiştir. İn vitro fertilizasyondan 18 saat sonra, fertilize edilmiş oositler in vitro kültür için sentetik ovidukt sıvısı (SOF) kültür medyumuna transfer edilmiştir. İki hücre, 8-hücre, morula ve blastosist sayıları kaydedilmiştir. FCS, SSS ve SR1 grupları için sırasıyla toplam 238, 243 ve 200 oosit kullanılmıştır. FCS, SSS ve SR1 gruplarında sırasıyla, 179 (%75,21), 177 (%72,84) ve 80 (%40,00); 121 (%50,84), 114 (%46,91) ve 38 (%19,00); 97 (%40,76), 98 (%40,03) ve 21 (%10,50); ve 71 (%29,83), 69 (%28,40) ve 10 (%5,00) oosit 2-hücre, 8-hücre, morula ve blastocyst aşamalarına gelişmiştir. SR1 grubunun kaydedilen tüm gelişim aşamalarındaki gelişim oranları FCS ve SSS gruplarının kaydedilen tüm gelişim oranlarından önemli düzeyde düşük bulunmuştur. FCS ve SSS grupları arasında gelişim oranları bakımından gelişim aşamalarının hiçbirinde fark bulunmamıştır. Sonuç olarak, SSS sığır oositlerinin in vitro maturasyonunda FCS’nin yerine kullanılabilen bir alternatifken, SR1 maturasyon medyumunda FCS’nin yerine kullanılabilecek iyi bir alternatif değildir.
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