Fertilization In vitro of follicular oocytes and cryopreservation of embryo fertilized and developed In vitro In Korean native cattle |
Sang-yong Choe1, Ill-keun Kong2, Young-kuk Joo2, Gyu-jin Rho1, Yong-kweon Kim3, Choong-saeng Park2 |
1College of Veterinary Medicine, Gyeongsang National University 2College of Agriculture, Gyeongsang National University 3Samcheonpo Office of Rural Guidance |
한우 난포란의 체외수정 및 체외수정란의 동결보존에 관한 연구 |
최상용1, 공일근2, 주영국2, 노규진1, 김용권3, 박충생2 |
1경상대학교 수의과대학 2경상대학교 농과대학 3삼천포 농촌지도소 |
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Abstract |
The ovaries of Korean Native cows or heifers were obtained from an abattoir and kept on 20 to $25^{circ}C$ and transported to laboratory within 2 hrs. The follicular oocystes were collected from 2~6mm follicles in diameter and classified into 3 grades by the morphology of cumulus cells attached. The oocytes were matured in vitro(IVM) for 24 hrs. in TCM-199 supplemented with $23{mu}g/ml$ FSH, $10{mu}g/ml$ LH, $1{mu}g/ml$ estradio-17 ${eta}$ and granulosa cells at $39^{circ}C$ under 5% $CO_2$ in air. They were fertilized in vitro(IVF) by incubation for 12 hrs. of epididymal spermatozoa pretreated with heparin, and then the zygotes were co-cultured in vitro(IVC) with oviductal epithelial cells for 7 to 9 days. Assessment of maturation revealed that 93.0%(147/158) of grade I oocytes had expanded of cumulus cells, which was higher(p<0.05) than the 79.4%(85/107) of grade II oocytes. Compared to epididymal sperm(32.9%), the insemination with frozen and thawed sperm resulted in slightly lower(20.5%), but not significant, development to morulae and blastocysts from grade I oocytes. Co-culture of bovine IVF embryos with oviductal epithelial cells improved the development to transferable embryos significantly(38.1%), compared to co-culture with granulosa cells(20.0%). When VF bovine embryos were vitrified at blastocyst, the post-thaw survival rate was obtained higher resulf for 1 min. equilibration time(82.6%) or 2 min.(73.9%) than 3 min.(18.2%) in EFS solution. |
Key Words:
In vitro fertilization, equilibration time, vitrification |
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