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KMID : 0359319930330040757
Korean Journal of Veterinary Research
1993 Volume.33 No. 4 p.757 ~ p.762
Fertilization In vitro of follicular oocytes and cryopreservation of embryo fertilized and developed In vitro in Korean native cattle






Abstract
The ovaries of Korean Native cows or heifers were obtained from an abattoir and kept on 20 to 25℃ and transported to laboratory within 2 hrs. The follicular oocystes were collected from 2∼6㎜ 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 ㎍/㎖ FSH, 10 ㎍/㎖ LH, 1 ㎍/㎖ estradio-17 β and granulosa cells at 39℃ under 5% CO₂ in air. They were fertilized in virto(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(38.196), compared to co-culture with granulosa cells(20.0%). When VF bovine embryps 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.
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학술진흥재단(KCI)