Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
권호 표지

Establishment of Freezing Conditions for Improving Cryosurvival in Miniature Pig Spermatozoa

미니돼지 정액의 동결-융해 후 생존율 향상을 위한 동결 조건 확립

  • Lee, Yong-Seung (College of Animal Life Sciences, Kangwon National University) ;
  • Yoo, Han-Jun (College of Animal Life Sciences, Kangwon National University) ;
  • Cheong, Hee-Tae (School of Veterinary Medicine, Kangwon National University) ;
  • Yang, Boo-Keun (College of Animal Life Sciences, Kangwon National University) ;
  • Woo, Jea-Seok (National Institute of Animal Science, Hanwoo Ex) ;
  • Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University)
  • 이용승 (강원대학교 동물생명과학대학) ;
  • 유한준 (강원대학교 동물생명과학대학) ;
  • 정희태 (강원대학교 수의과대학) ;
  • 양부근 (강원대학교 동물생명과학대학) ;
  • 우제석 (국립축산과학원) ;
  • 박춘근 (강원대학교 동물생명과학대학)
  • Received : 2011.09.02
  • Accepted : 2011.09.09
  • Published : 2011.09.30
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Abstract

This study was conducted to establish a freezing method of miniature pig spermatozoa. The semen 더aculated from PWG M-type miniature pig was collected by gloved-hand method. The semen was diluted with same volume extender (m-Modena B). The frozen solution used frozen solution of four different (LEY, TCG, BF-5 and m-Modena+egg yolk) for find optimal frozen solution in miniature pig sperm. The diluted semen for frozen rate assay was added to LEY solution (solution I: 11% lactose+egg yolk; solution II: solution I+glycerol+OEP), and frozen depending on freezing rate by the three different freezing methods (A: until $5^{\circ}C$ for 1 hrs, holding at $-102^{\circ}C$ for 10 min; B: until $5^{\circ}C$ for 2 hrs, holding at $-102^{\circ}C$ for 10 min; C: until $5^{\circ}C$ for 3 hrs, holding at -80 and $-102^{\circ}C$ for 10 min). Semen cooled until $5^{\circ}C$ was added with glycerol 1, 3 and 5%, and take a equilibrium time for 0, 10 and 30min. Frozen-thawed sperm were evaluated for viability, acrosomal status and morphological abnormality. The results of frozen-thawed sperm ability by frozen solution, viability was higher in LEY solution compared to other three different frozen solution. AR pattern of LEY solution were lower than other three different frozen solution. The results of freezing rate, viability was higher in B method compared to other methods (p<0.05). Acrosomal statute was intacted in A and B methods than C method. The experiment for glycerol condition was showed that sperm viability was higher in extender with 1% and 3% glycerol and equilibrium time of 0 min. The acrosome damage was lower in extender with 1% glycerol and equilibrium time of 10 min than other conditions. In conclusion, the optimal conditions for cryopreservation of miniature pig spermatozoa obtained in LEY frozen solution, cooling rate of 1~2 hrs, 1~3% glycerol concentrations and glycerol equilibrium time of 0~10 min.

Keywords

References

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