Ahmet ESER, EBRU GÜREL GÜREVİN, Hatem ATALLA, KEMAL AK, SEMA BİRLER, SERHAT PABUCCUOĞLU, AYÇA ÜVEZ, ELİF İLKAY ARMUTAK, RAMAZAN ARICI, KAMBER DEMİR, SELİN YAĞCIOĞLU, Nur ERSOY, Nihan COŞKUN, MİTHAT EVECEN

Effects of Ovary Transport and Storage Temperature on In Vitro Maturation and Cumulus Cell Apoptosis Rates in Cat Oocytes

Çalışmanın amacı, ovaryumların iki farklı sıcaklıkta (37°C ve 4°C) taşınma ve soğukta 24 saat bekletilmenin kumulus hücrelerindeki apoptoza ve kedi oositlerinin in vitro olgunlaşma oranları (IVM) üzerine etkilerini incelemektir. Kısırlaştırılmış 15 kediden ovaryumlar alındı ve yarısı 37°C, diğer yarısı da 4°C'de olmak üzere, fosfat tampon tuzlu solüsyonunda (PBS) laboratuara taşındı. Soğukta bekletmenin etkilerini belirlemek içinse, 4°C 'de taşınan ovaryumların yarısı, aynı sıcaklıkta olmak üzere 24 saat bekletildi. Seçilen kumulus oosit kompleksleri (COCs), %100'e yakın nemin sağlandığı %5 CO2'li inkübatörde mineral yağ altındaki 500 ?L modifiye Sentetik Ovidukt Medyumu (mSOF) içeren dört gözlü petrilerde olmak üzere 38°C'de 48 saat süreyle olgunlaştırıldı. Apoptozun etkileri, hem iki farklı taşıma grubunda, hem de soğukta 24 saat bekletilen grupta olmak üzere in vitro olgunlaşmanın hemen öncesinde kumulus hücrelerinde test edildi. Apoptosis derecesi, deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) test ile ölçüldü. Oositlerin in vitro olgunlaşma düzeyleri Hoechst (33342) boyama ile belirlendi. Apoptotik hücre oranları, 37°C ve 4°C taşıma gruplarında sırasıyla; %19.40 ve %21.55 olarak belirlendi (P>0.001). Aynı değer, 24 saatlik soğukta bekletilen grupta ise daha yoğun olarak (%34.80) görüldü (P0.05) iken, 24 saat soğukta bekletme grubunda ise %18.90 olarak bulundu (P

Kedi Oositlerinin In Vitro Olgunlaştırılması ve Kumulus Hücrelerinin Apoptoz Oranları Üzerine, Ovaryum Taşıma ve Saklama Sıcaklığının Etkisi

The objective of the present study was to examine the effects of two different transport temperature (37°C vs 4°C) and cold storage of ovaries for 24 h on cumulus cell apoptosis and maturation rates of cat oocytes in vitro. Ovaries were collected from 15 ovariohysterectomized domestic cats and maintained and transported to the laboratory in phosphate buffer saline at 37°C and 4°C. In order to determine the effects of storing time, some ovaries transported at 4°C were stored at the same temperature for 24 h. Selected cumulus oocyte complexes (COCs) were matured for 48 h at 38°C in four-well petri dishes containing 500 μL of modified oviduct medium (mSOF) under mineral oil in a 5% CO2 incubator with nearly 100% humidified. The morphological features of apoptosis were analysed in the cumulus cells at the beginning of in vitro maturation in both transporting temperature groups and after 24 h of cold stored group. The degree of apoptosis in cumulus cells were measured by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL). The IVM rates of oocytes were determined using Hoechst (33342) staining. Although the apoptotic morphological features were seen rarely and in similar rates in 37 and 4°C transporting groups (19.40 and 21.55%, P>0.001), it was seen more intensely in the 24 h cold stored group (34.80%, P0.05), and importantly lower at 4°C transporting and 24 h cold stored groups (18.90%, P

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