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KMID : 0900919970210020103
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Korean journal of Animal Reproduction
1997 Volume.21 No. 2 p.103 ~ p.109
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Studies on the Development of Easy Cryopreservation Technique of Bovine Embryos II. Effects of Equilibration of Cryoprotectants, Temperature and Time of Thawing and 1 Step Straw Method on In Vitro Developmental Rates of Embryos
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Kim Sang-Keun
Hyun Byung-Hwa
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Abstract
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The studies on the carried out to investigate to determine the optimum thawing temperature and equilibration time and 1 step straw method of frozen bovine embryos. The follicular oocytes were cultured in TCM-199 medium containing 10 IU/ml PMSG(Sigma, USA), 10 IU/ml hCG(Sigma, USA), 1 mutextrm{g}/ml beta-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% CO2 in air at 38.5^{circ}C and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation of heparin. The bovine embryos following dehydration by cryoprotective agents and various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 30^{circ}C water. Survival and in vitro developmental rate was defined as developmental rate on in vitro culture or FDA-test. The results are summarized as followes : 1. The equilibration time on in vitro developmental rates of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (10~20 min.). 2. The temperature thawed at 30^{circ}C after rapid freezing of bovine embryos resulted in a significantly higher in vitro developmental rate than did at 20^{circ}C and 35^{circ}C. 3. The thawing time on in vitro developmental rates of bovine embryos was attained after short period of time(1~5 min.) in the freezing mediuim higher than long period of time(10min.). 4. The in vitro developmental rates of bovine embryos after rapid frozen-thawing by 1 step straw method in the freezing medium added 1.5M, 2.0M glycerol, DMSO, propanediol and 0.25M, 0.50M, 0.75M, 1.00M sucrose were 12.5~19.4%, 10.0~15.6%, 9.1~13.8% and 6.7~12.9%, respectively.
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KEYWORD
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Bovine embryos, Equilibraion, Rapid freezing, Thawing, 1 step straw method
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