Korean Journal of Veterinary Research 1993;33(4):757-762.
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
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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