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KMID : 1005220120270040253
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Journal of Embryo Transfer
2012 Volume.27 No. 4 p.253 ~ p.258
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Cryopreservation of Embryo by Concentration of Ethylene Glycol and Day 6, 7, 8, 9 Embryo in Korean Native Cattle (Hanwoo)
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Park Joung-Jun
Yoo Han-Jun
Choi Hye-Won
Cheong Ki-Soo
Kim Ji-Tae
Park Choon-Keun
Yang Boh-Suk
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Abstract
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This study was carried out to effects of ethylene glycol concentration, sucrose and culture day of in vitro production embryo on slow-down freezing in Hanwoo. 6, 7, 8 and 9 day embryos produced in vitro were frozen using 1.8M EG+0.1M sucrose, 1.8M EG+0.5% BSA and 1.5M EG+0.1M sucrose media. Survivability was confirmed after frozenthawed 24 and 48h and ICM, TE cell number were counted by Hoechst 33342 and PI staining after frozen-thawed 24h. As a result, 1.8M EG+0.1M sucrose group was most significantly (p<0.05) higher compared with the other treatment groups on survivability, TE and total cell number after frozen-thawed 24h (94.2 ± 2.6%, 94.67 ± 3.4 and 129.67 ± 5.5). ICM number did not found significant (p<0.05) differences between the three treatment groups. in 6, 7, 8 and 9 day of embryos using three types of freezing media, frozen-thawed, 1.8M EG+0.1M sucrose groups with embryos cultured 8 day was significantly (p<0.05) highest survivability to 98.3 ± 1.7% after frozen-thawed 24h. 1.5M EG+0.1 sucrose group with embryos cultured 9 day was significantly higher survivability than group of embryos cultured 8 day after frozen-thawed 24 and 48h. In conclusion, 1.8M EG+0.1M sucrose media is considered to be effective to cryopreservation of embryos cultured 8 and 9 day.
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KEYWORD
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cryopreservation , ethylene glycol , embryo stage , Hanwoo
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