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In Vitro Development of Porcine Oocytes Following Intracytoplasmic Injection of Freeze-Dried Spermatozoa with Trehalose

Trehalose에 의하여 동결 건조된 정자의 돼지 난자 내 직접주입 후 체외 배발달

  • Kang, Hwa-Hyung (Department of Animal Science, College of Agriculture and Life Sciences, Chonnam National University) ;
  • Lee, Ji-Woong (Department of Animal Science, College of Agriculture and Life Sciences, Chonnam National University) ;
  • Kang, Man-Jong (Department of Animal Science, College of Agriculture and Life Sciences, Chonnam National University) ;
  • Kim, Kwang-Hyun (Department of Animal Science, College of Agriculture and Life Sciences, Chonnam National University) ;
  • Moon, Seung-Ju (Department of Animal Science, College of Agriculture and Life Sciences, Chonnam National University)
  • 강화형 (전남대학교 동물자원학부) ;
  • 이지웅 (전남대학교 동물자원학부) ;
  • 강만종 (전남대학교 동물자원학부) ;
  • 김광현 (전남대학교 동물자원학부) ;
  • 문승주 (전남대학교 동물자원학부)
  • Received : 2014.01.07
  • Accepted : 2014.03.25
  • Published : 2014.03.31

Abstract

The objectives of this study were to investigate the effects of trehalose as a cryoprotectant for porcine freeze-dried spermatozoa, to find the optimal freeze-drying time and storage periods of freeze-dried spermatozoa, and to find out pronuclear formation rates, cleaved rates, and embryo development through intracytoplasmic injection of freeze-dried spermatozoa on porcine oocytes. The survival rates of spermatozoa after freeze-drying with trehalose treatment were significantly higher than those of them without trehalose treatment (p<0.05). The highest survival rates were found at 75 mM trehalose treatment. The longer storage periods after freeze-drying seemed to have a lower survival rates. Development in culture of pig by ICSI with trehalose treatment were significantly higher than those of them without trehalose treatment (p<0.05). Shorter freeze-drying time of spermatozoa was resulted in the highest cleaved rates and embryo development.

본 연구는 돼지 정자의 동결 건조 시 동결 보호제인 trehalose의 효과와 동결 건조 시간과 동결 건조 후 저장 기간에 따라 정자의 생존성과 체외 성숙 난자 내 동결 건조 정자를 직접 주입한 후 전핵 형성율, 난할율 그리고 배발달 성적을 조사하였다. 동결 건조 후 정자의 생존율은 trehalose 무첨가구에 비해 trehalose를 첨가한 처리구에서 높은 생존율을 보였으며, 75 mM의 trehalose를 첨가하여 동결 건조한 정자들의 생존율이 가장 높은 것으로 나타났다. 또한, 동결 건조 후 저장 기간이 길어질수록 생존율이 낮아지는 경향이었다. 체외 성숙 난자 내 동결 건조 정자를 직접 주입 후 전핵 형성율은 trehalose 첨가구에서 유의적으로 높았으며(p<0.05), 난할율과 배발달 성적도 trehalose 첨가구에서 유의적으로 높았고(p<0.05), 정자의 동결 건조 시간이 짧을수록 높은 난할율과 배발달율을 보였다.

Keywords

References

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