The effects of various protein supplementations on In vitro maturation of cat oocytes

Bu çalışma, medyuma katılan değişik protein kaynaklarının kedi oositlerinin in vitro olgunlaştırılması üzerine etkilerini araştırmak amacıyla yapıldı. Çalışmada, kısırlaştırma operasyonu uygulanmış 40 kediden kazanılan 1013 oosit kullanıldı. Ovaryumlar, 2530° C sıcaklıktaki PBS solüsyonu içerisinde laboratuvara taşındı. Kazanılan oositler, farklı protein kaynaklarını içeren Ham’s F-10 medyumu içerisinde ve 38.5°C sıcaklık, %100’e yakın nem ve gaz karışımının (%5 O2, %5 CO2, %90 N2) sağlandığı inkübatör ortamında 48 saat süreyle olgunlaşmaya bırakıldı. Sunulan çalışmada dört uygulama grubu oluşturuldu. Grup I: Fetal Buzağı Serumu (FCS), Grup II: Sığır Serum Albumini (BSA), Grup III: Östrustaki Kedi Serumu (OCS), Grup IV: Kontrol (protein katkısı yok). Olgunlaşma sürecinin sonunda, oositler fikse edilerek boyandı. Olgunlaşma oranları, faz - kontrast mikroskopta (x400) yapılan incelemelerle belirlendi. Metafaz II (M II) dönemine ulaşma oranları Grup I, II, III ve IV için sırasıyla %6.2 (14/255), %16.5 (43/260), %13.1 (36/273) ve %13.3 (30/225) olarak bulundu. Protein kaynağı olarak FCS’nin kullanıldığı Grup I deki olgunlaşma oranları, diğerlerinden oldukça düşük olarak bulundu (P

Anahtar Kelimeler:

oosit, kedi, in vitro, oosit olgunlaşması, yumurtalıklar, protein kaynakları, protein zenginleştiriciler

Kedi oositlerinin in vitro olgunlaştırılması üzerine değişik protein katkılarının etkileri

This study has been conducted to evaluate the effects of certain media containing various protein sources on the in vitromaturation (IVM) of cat oocytes. In this study 1013 oocytes obtained from 40 cats underwent ovario-hysterectomy surgeriesfor the purposes of spaying were used. Ovaries were brought to the laboratory in Dulbecco’s PBS at 25-30°C. The recoveredoocytes were left to mature for 48 hours in Ham’s F-10 media supplemented with different protein sources. In this study fourexperimental groups were designed: Group I: Fetal Calf Serum (FCS), Group II: Bovine Serum Albumin (BSA), Group III: EstrusCat Serum (OCS), Group IV: Control (No protein supplementation). The incubator conditions were 38.5°C temperature, almost100% humidity and a gaseous mixture (5% O2, 5% CO2, 90% N2). At the end of the maturation period, the oocytes were fixed and stained. The maturation rates were determined under 400 x magnification on a phase-contrast microscope. Metaphase II(M II) rates were 6.2% (14/255), 16.5% (43/260), 13.1% (36/273) and 13.3% (30/225) in Groups I, II, III and IV, respectively. The maturation rate in the Group I where FCS was used as protein additive was significantly lower (P<0.001) than other groups.The ratio of oocytes reaching M I+M II stages was 28.6% (73/255), 40.3% (105/260), 31.1% (85/273) and 30.2% (68/225),respectively. The difference in the Group II (BSA added), was significantly superior to the other groups (P<0.05). In conclusion,although homologous protein source OCS was ineffective and FCS has negative effects, it was determined that using BSA asthe protein additive for the medium will be beneficial for in vitro maturation of cat oocytes.

Keywords:

oocytes, cats, in vitro, oocyte maturation, ovaries, protein sources, protein supplements,

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