Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Establishment of the Convenient Boar Semen Freezing Method and Assessment of Viability in Frozen/Thawed Boar Semen

돼지 정액의 간편 동결 방법 확립과 동결 정액의 융해 후 생존성 평가

  • Kim Seong-Kon (College of Animal Life Sciences, Kangwon National University) ;
  • Jang Hyun-Yong (College of Animal Life Sciences, Kangwon National University) ;
  • Park Dong-Heon (College of Animal Life Sciences, Kangwon National University) ;
  • Park Chun-Keun (College of Animal Life Sciences, Kangwon National University) ;
  • Cheong Hee-Tae (School of Veterinary Medicine, Kangwon National University) ;
  • Kim Choung-Ik (College of Animal Life Sciences, Kangwon National University) ;
  • Yang Boo-Keun (College of Animal Life Sciences, Kangwon National University)
  • 김성곤 (강원대학교 동물생명과학대학) ;
  • 장현용 (강원대학교 동물생명과학대학) ;
  • 박동헌 (강원대학교 동물생명과학대학) ;
  • 박춘근 (강원대학교 동물생명과학대학) ;
  • 정희태 (강원대학교 수의학부대학) ;
  • 김정익 (강원대학교 동물생명과학대학) ;
  • 양부근 (강원대학교 동물생명과학대학)
  • Published : 2006.03.01
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Abstract

This study was conducted to establish a convenient freezing method of boar semen. Boar semen was cooled until $5^{\circ}C$ for 3 hrs using cell freezer and loaded into straws. Semen straws were frozen in different steps in strofoam box filled with $LN_2$. Highest sperm viability (54.0%) was obtained by 1-step freezing(holding at 10 cm height from the surface of $LN_2$ for 10 min). Sperm viability increased by holding at $-102^{\circ}C$ for 10min (74.0%, P<0.05). In thawing regime, sperm viability was significantly higher in $37^{\circ}C$ group than in $52^{\circ}C$ group. The sperm characteristics did not differ between 1-step and 3-step. After IVF using frozen-thawed boar semen, developmental rate of embryos to the morula+blastocyst stage was in 1-step freezing group than that of 3-step freezing group (27.5 vs 14.7%, P<0.05). The result shows that the 1-step freezing with holding at $-102^{\circ}C$ for 10min before plunging into $LN_2$ is a convenient and easy freezing method for boar semen.

본 연구는 돼지정액의 보다 간편하고 손쉽게 동결시킬 수 있은 방법을 확립하기 위하여 수행하였다. 돼지정액은 3시간에 걸쳐 $5^{\circ}C$까지 냉각 후 Straw에 봉입하고 다양한 방법 및 step에 의해 스티로폼 용기 내에 들어있는 $LN_2$ 중에서 동결하였다. 정자의 생존성은 $LN_2$ 표면으로부터 10 cm 위에서 10분간 정치 후 침적할 경우 가장 높게 나타났다(54.0%). Straw를 $-102^{\circ}C$에서 10분간 정치시킨 처리구가 여타구보다 높은 생존성이 얻어졌다(74.0%, P<0.05). 응해 방법에 따른 동결정액의 생존성 실험에서는 $37^{\circ}C$의 융해구가 $52^{\circ}C$ 융해구보다 유의적으로 높은 결과를 나타냈다(P<0.05). 1단계 동결 방법과 3단계 동결방법으로 돼지 정액을 동결시킨 결과 정액의 일반적 특성 및 첨체의 이상 유무를 평가하는 CTC 검사에서는 커다란 차이가 인정되지 않았다. 동결정액을 이용한 체외수정 결과에서는 상실배기 이상 발육율에서 1-step이 3-step보다 높은 발육율을 나타내었다(27.5 vs 14.7%, P<0.05). 본 실험의 결과 돼지정액 동결 시 $-102^{\circ}C$에서 10분간 정치시키는 1단계 동결방법이 간편하면서 유리한 동결 방법으로 활용될 수 있음을 보여준다.

Keywords

References

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