Development of In Vitro Derived Sheep Embryos to the Blastocyst Stage

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

Pages : 891-894

View : 15 | Download : 5

Publication Date : 0000-00-00

Article Type : Research Paper

Abstract :The objective of the present study was to mature and fertilize primary sheep oocytes in vitro and to develop them to the blastocyst stage. Ovaries from slaughtered Kıvırcık ewes were used. The ovaries were transported to the laboratory in a vacuum flask in which 30-35 °C PBS insert ignore into journalissuearticles values(phosphate buffered saline); was included. The cumulus-oocyte complexes were collected by rupturing and washing the follicle wall with oocyte washing medium. They were then selected under a stereo microscope and maturated for 23-24 hours 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);, under 5% CO2 atmosphere and at 38.5 °C temperature. Semen collected and pooled from three Kivircik rams by electro-ejaculation and prepared for in vitro fertilization by Percoll-gradient method was incubated for 20-21 hours with oocytes in BSOF medium supplemented with 2% sheep estrous serum insert ignore into journalissuearticles values(SES); insert ignore into journalissuearticles values(0.8x106 spermatozoon/ml);. Presumptive zygotes were cultured in SOF medium for 8 days in synthetic oviduct medium insert ignore into journalissuearticles values(SOF); under an atmosphere of 5% CO2, 5% O2 and 90% N2. Glucose insert ignore into journalissuearticles values(1.5mM); was added to the culture medium on Day 4. Out of 171 oocytes, 151 cleaved insert ignore into journalissuearticles values(88.3% );. Fifty-six morulae insert ignore into journalissuearticles values(37.1% ); and 14 blastocysts insert ignore into journalissuearticles values(9.3% ); were obtained from these in vitro derived embryos.
Keywords : Sheep, in vitro fertilization, in vitro culture, development to blastocyst stage