PREGNANCY RATE AND SURVIVAL IN CULTURE OF IN VITRO FERTILIZED BOVINE EMBRYOS FROZEN IN VARIOUS CRYYOPROTECTANTS AND THAWED USING A ONE-STEP SYSTEM

Suzuki, T.; Takagi, M.; Yamamoto, M.; Boediono, A.; Saha, S.; Sakakibara, H.; Oe, M.

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자료유형: 학술대회자료

저자정보: Suzuki, T. (United Graduate School of Veterinary Science Yamaguchi University) Takagi, M. (United Graduate School of Veterinary Science Yamaguchi University) Yamamoto, M. (United Graduate School of Veterinary Science Yamaguchi University) Boediono, A. (United Graduate School of Veterinary Science Yamaguchi University) Saha, S. (United Graduate School of Veterinary Science Yamaguchi University) Sakakibara, H. (United Graduate School of Veterinary Science Yamaguchi University) Oe, M. (United Graduate School of Veterinary Science Yamaguchi University)

저널정보: 한국동물번식학회 한국수정란이식학회 학술대회 한국수정란이식학회 1997년도 국제학술대회 및 Workshop

발행연도: 1997.1

수록면: 27 - 34 (8page)

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Bovine oocytes surrounded with compact cumulus cells were cultured for 20 to 22 hours($38^{\circ}C$, 5% $CO_2$) in modified TCM-199 medium supplemented with 5% superovulated cow serum(SCS) and inseminated by in vitro capacitated spermatozoa. Day 7 to 8 embryos were equilibrated for 10 minutes in 1.3M methyl cellosolve(MC) <1.1M diethylene glycol(DEG), 1.8M ethylene glycol(EG), 1.6M propylene glycol(PG) and 1.1 M 1,3-butylene glycol(BG) solutions. They were then loaded into 0.25ml straws, placed into an alcohol bath freezer at $0^{\circ}C$, cooled from $0^{\circ}C$ to $-6^{\circ}C$ at $-1^{\circ}C$/minute, seeded, held for 10 minutes, and stored in liquid nitrogen. After thawing in $30^{\circ}C$ water, the embryos wee rehydrated in TCM-199 medium and then cultured for 48 hours in TCM-199 plus 5% SCS. Embryos were considered viable if they progressed to later developmental stages with a good morphology. Some of the embryos frozen in each cryoprotectant were thawed and transferred non-surgically without removing the cryoprotectant. Hatched embryos survived freezing and one-step dilution as follows : EG(50.0%), MC(53.6%), DEG(56.9%), PG(58.0%) and BG(11.5%). The survival rate of embryos cooled at -0.3^{\circ}C$ vs. $-0.5^{\circ}C$/minute was not significantly different(P<0.05), however, blastocysts hatched most often (P<0.01) in vitro when cooled at a rate of $0.3^{\circ}C$/minute(64.6%), 31/48) than at $-0.5^{\circ}C$/minute(22.6%, 12/53). Pregnancy rates resulting from embryos frozen in the different cryoprotectants were as follows : MC(48%, 10/21); DEG(30%, 3/10); EG(74%, 20/27); and PG(40%, 4/10). These results indicate that MC, DEG, EG and PG have utility as cryoprotectants for the freezing and thawing of IVF Bovine embryos.

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