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


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.


  1. Ahmad K (1984): Effect of thaw rates on survival of buffalo spennatozoa frozen straws. J Dairy Sci 67:1535-1538
  2. Almlid T, Hofmo PO (1996): Brief review of frozen semen application under Norwegian AI service conditions. Reprod Dom Anim 31:169-173
  3. Bwanga, CO (1991):. Cryopreservation of boar semen. I : A literature review. Acta Vet Scand 32: 431-453
  4. Cordova A, Ducolomb Y, Jimenez I, Cassas E, Bonilla E, Betrancourt M (1997): In vitro fertilizing capacity of frozen-thawed boar semen. Theriogenology 47:1309-1317
  5. Eriksson BM, Petersson H, Rodriguez-Martinez H (2002): Field fertility with exported boar semen frozen: in .the new FlacPack container. Theriogenology 58:1065-1079
  6. Fiser PS, Fairfull RW (1990): Combined effect of .glycerol concentration and cooling velocity on motility ad acrosomal integrity of boar spennatozoa frozen in 0.5 ml straw. Mol Reprod Dev 25:123-129
  7. Fiser PS, Rairfull RW, Marcus GJ (1986): The effect of thawing velocity on survival and acrosomal integrity of ram spennatozoa frozen at optimal and suboptimal rates in straws. Cryobiology 23:141-149
  8. Hammersted RH, Graham JK, Nolan JP (1990): Cryopreservation of mammalian sperm : What we ask them to survive. J Androl 11:73-88
  9. Huang SY, Kuo YH, Lee WC, Tsou HL, Lee YP, Chang HL, Li-Jun Huo JJ, Ma XH, Yang ZM (2002): Assessment of spenn viability, mitochondrial activity, capacitation and acrosome intactness in extended boar semen during long-tenn storage. Theriogenology 58:1349-1360
  10. Kumar S, Millar JD, Waston PF (2003): The effect of cooling rate on the survival of cryopreserved bull, ram, and boar spennatozoa : A comparison of two controlled-rate cooling machines. Cryobiology 46: 246-253
  11. Maxwell WMC, Johnson LA (1997): Membrane status of boar spennatozoa after cooling or cryopreservation. Theriogenology 45:209-219
  12. Medrano A, Watson PF, Holt WV (2002): Importance of cooling rate and animal variability for boar spenn cryopreservation : Insights from the cryomicroscope. Reproduction 123:315-322
  13. Nagai T, Takahashi T, Masuda H, Shioyo Y, Kumayama M, Fukushima M, Iwasaki S, Hanada A (1988): In vitro fertilization of pig oocytes by frozen boar spennatozoa. J Reprod Fertil 84:585-591
  14. Neild DM, Gadella BM, Maria GC, Marcelo H, Miragaya C, Colenbrander B, Aguero A (2003): Membrane changes during different stages of a freeze-thaw protocol for equine semen cryopreservation. Theriogenology 59:1693-1705
  15. Salisbury GW, VanDemark NL, Lodge JR (1978): Physiology of reproduction and artificial insemination of cattle. W. H. Freeman and Co. 2nd (eds), San Francisco, USA
  16. Watson PF (1996): Cooling of spennatozoa and fertilizing capacity. In Rath D, Johnson LA and Weitze KF (eds), boar semen preservation III. Proc. 3rd Int. Cont. Boar semen preservation, Mariencee, Gennany, August, 1995. Reprod Domest Anim Vol 31 Blackwell, Berlin, pp. 135-140
  17. Zheng YS, Fiser P, Sirard MA (1992): The use of ejaculated boar semen after freezing in 2 or 6% glycerol for in vitro fertilization of porcine oocytes matured in vitro. Theriogenology 38:1065-1075
  18. 加藤碇史良, 井上揚一, 贋野森, 入浴明, 西川義正 (1976): Effect of thawing procedure on motility and acrosome system of frozen boar spermatozoa. 日本凍結精液硏究學會報48:15-19
  19. 이광원, 지설하, 손동수, 김학규, 박창식, 김친철, 최진성, 고문석, 정행기, 김현 (1989): 5 m1 straw에 보존한 돼지 희석정 액의 생존성과 수정능력에 관한 연구. 한국동물자원과학회지 31:158-161
  20. 임종우, 윤창현 (1973): 돈정액 보존에 관한 연구. 축산진흥연구소보 1:96-102
  21. 정영호, 서경덕, 김광식, 심금섭, 이장희 (1999): 동결 보존한 돼지정액의 융해조건이 정자의 생존율과 첨체변화에 미치는 효과. 한국수정란이식학회지 14:131 -137
  22. 최영진, 박춘근, 정희태, 김정익, 박동헌, 장현용, 양부근 (2002): 항산화제와 Growth factors 첨가가 돼지 체외 수정란의 체외발육에 미치는 효과. Korean J Anim Reprod 26:143-151