Transfer of In Vitro Produced Sheep Embryos

Authors : Sema BİRLER, Serhat PABUÇÇUOĞLU, Hatem ATALLA, Serhat ALKAN, Özen Banu ÖZDAŞ

Pages : 1421-1426

View : 17 | Download : 6

Publication Date : 0000-00-00

Article Type : Research Paper

Abstract :The objective of the present study was to transfer sheep embryos produced in vitro to recipient ewes. Ovaries were taken from slaughtered Kıvırcık ewes and transferred to the laboratory in phosphate buffered saline insert ignore into journalissuearticles values(PBS); at 30-35 ºC. The cumulus-oocyte complexes were obtained by slicing and washing 1-6 mm diameter follicles and matured for 24 h in medium 199 supplemented with sodium pyruvate, follicle stimulating hormone insert ignore into journalissuearticles values(FSH);, luteinizing hormone insert ignore into journalissuearticles values(LH); and 10% fetal calf serum insert ignore into journalissuearticles values(FCS); at 38.5 ºC under 5% CO2 in humidified atmosphere. Fresh semen was collected from three Kıvırcık rams, pooled and prepared for in vitro fertilization by the percoll-gradient method. Matured oocytes were transferred into synthetic oviduct fluid insert ignore into journalissuearticles values(SOF); based fertilization medium supplemented with 2% sheep oestrous serum insert ignore into journalissuearticles values(SES); and co-incubated with semen insert ignore into journalissuearticles values(0.8 x 106 spermatozoon/ml); for 20-21 h. After fertilization, presumptive zygotes were transferred into SOF medium and incubated for 8 days under an atmosphere of 5% CO2, 5% O2 and 90% N2 at 38.5 ºC. Glucose insert ignore into journalissuearticles values(1.5 mM); was added to the culture medium on day 4. In culture, embryos were checked for cleavage and embryo development on days 4 and 8, respectively. A total of 534 oocytes were inseminated in vitro, 407 insert ignore into journalissuearticles values(76.2%); cleaved, and 119 insert ignore into journalissuearticles values(29.2%); and 69 insert ignore into journalissuearticles values(16.9%); reached the morula and blastocyst stages, respectively. Forty days after the transfer of 40 blastocysts into the uteri of 17 recipient ewes, pregnancy was detected in 10 ewes insert ignore into journalissuearticles values(58.8%); by ultrasonography and 7 ewes gave birth to 8 live lambs. The other 3 ewes, which had 7 fetuses, could not have live offspring. According to the transferred embryos, the implantation rate was 37.5% and the live lamb rate was 20.0%.
Keywords : Sheep, in vitro production, embryo transfer