Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Effect of Various Percoll Washings on Motile Sperm Recovery Rate and Motion Kinematics in Frozen-thawed Bovine Semen

다양한 Percoll 세척 방법이 동결-융해된 한우 정자의 회수율 및 운동역학에 미치는 영향

  • Yoon, Sung-Jae (Department of Animal Science and Technology, School of Bioresource & Bioscience, Chung-Ang University) ;
  • Park, Yoo-Jin (Department of Animal Science and Technology, School of Bioresource & Bioscience, Chung-Ang University) ;
  • Cheong, Jin-Yong (Department of Animal Science and Technology, School of Bioresource & Bioscience, Chung-Ang University) ;
  • Jeong, Kyu-Hyun (Department of Animal Science and Technology, School of Bioresource & Bioscience, Chung-Ang University) ;
  • Kim, Min-Seop (Department of Animal Science and Technology, School of Bioresource & Bioscience, Chung-Ang University) ;
  • Yoo, Sae-Mi (Department of Animal Science and Technology, School of Bioresource & Bioscience, Chung-Ang University) ;
  • Kim, Yun-Hee (Department of Animal Science and Technology, School of Bioresource & Bioscience, Chung-Ang University) ;
  • Kwon, Woo-Sung (Department of Animal Science and Technology, School of Bioresource & Bioscience, Chung-Ang University) ;
  • Mohamed, El-Sayed A. (Department of Animal Science and Technology, School of Bioresource & Bioscience, Chung-Ang University) ;
  • Pang, Myung-Geol (Department of Animal Science and Technology, School of Bioresource & Bioscience, Chung-Ang University)
  • 윤성재 (중앙대학교 생명자원공학부 동물생명공학) ;
  • 박유진 (중앙대학교 생명자원공학부 동물생명공학) ;
  • 정진용 (중앙대학교 생명자원공학부 동물생명공학) ;
  • 정규현 (중앙대학교 생명자원공학부 동물생명공학) ;
  • 김민섭 (중앙대학교 생명자원공학부 동물생명공학) ;
  • 유새미 (중앙대학교 생명자원공학부 동물생명공학) ;
  • 김연희 (중앙대학교 생명자원공학부 동물생명공학) ;
  • 권우성 (중앙대학교 생명자원공학부 동물생명공학) ;
  • ;
  • 방명걸 (중앙대학교 생명자원공학부 동물생명공학)
  • Received : 2011.02.25
  • Accepted : 2011.03.03
  • Published : 2011.03.31
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Abstract

The objective of this study was to examine the effect of various discontinuous Percoll washing conditions on motile sperm recovery rate and motion kinematics. Frozen semen samples from 3 bulls (0.5 ml plastic straws, 6% glycerol in egg yolk-Tris-glycerol extender) were thawed in $37^{\circ}C$ water bath for 1 min. After thawing, the mixed semen samples were randomly allocated to 12 treatment groups. Briefly, the spermatozoa were centrifuged for three different time lengths (10, 20, and 30 min) at two gravities ($300{\times}g$ and $700{\times}g$) through two concentrations of discontinuous Percoll density gradient of 1 ml 90%: 1 ml 45% Percoll and 2 ml 90%: 2 ml 45% Percoll to remove extender, debris, and dead spermatozoa. Motile sperm recovery rate and motion kinematics were evaluated by computer assisted sperm analyzer using Makler counting chamber. Sperm motility (%) and motile sperm recovery rate showed similar pattern in all treatment groups. However, sperm motility (%) and motile sperm recovery rate were highest at $700{\times}g$ for 30 min through a discontionous Percoll density gradient of 1 ml 90%: 1 ml 45% Percoll. There were no significant differences in motion kinematics after various Percoll washings. These results suggest that force of centrifugation, centrifugation time, and Percoll volume significantly affect motile sperm recovery rate.

Keywords

References

  1. Aitken RJ, Clarkson JS (1988): Significance of reactive oxygen species and antioxidants in defining the efficacy of sperm preparation techniques. J Androl 9:367-376.
  2. Anzar M, Graham EF, Iqbal N (1997): Post-thaw plasma membrane integrity of bull spermatozoa separated with a Sephadex ion-exchange column. Theriogenology 47:845-856. https://doi.org/10.1016/S0093-691X(97)00040-X
  3. Henkel RR, Schill WB (2003): Sperm preparation for ART. Reprod BioI Endocrinol 1:108. https://doi.org/10.1186/1477-7827-1-108
  4. Holt C, Holt WV, Moore HD, Reed HC, Curnock RM (1997): Objectively measured boar sperm motility parameters correlate with the outcomes of onfarm inseminations: results of two fertility trials. J Androl 18:312-323.
  5. Kim YJ, Ji DB, Oh HG (2000): Studies on HOS test and CTC test for viability and capacitation of frozen-thawed canine sperm. Korean J Vet Clin Med 17:431-437.
  6. Oh SA, You YA, Park YJ, Pang MG (2010): The sperm penetration assay predicts the litter size in pigs. Int J Androl 33: 604-612.
  7. Oshio S (1988): Apparent densities of spermatozoa of various mammalian species. Gamete Res 20:159-164. https://doi.org/10.1002/mrd.1120200206
  8. Parrish JJ, Krogenaes A, Susko-Parrish JL (1995): Effect of bovine sperm separation by either swim-up or Percoll method on success of in vitro fertilization and early embryonic development. Theriogenology 44:859-869. https://doi.org/10.1016/0093-691X(95)00271-9
  9. Parrish JJ, Susko-Parrish JL, Leibfried-Rutledge ML, Critser ES, Eyestone WH, First NL (1986): Bovine in vitro fertilization with frozen-thawed semen. Theriogenology 25:591-600. https://doi.org/10.1016/0093-691X(86)90143-3
  10. Reddy JM, Stark RA, Zaneveld LJD (1979): A higher molecular weight antifertility factor from human seminal plasma. J Reprod Fertil 57:437-446. https://doi.org/10.1530/jrf.0.0570437
  11. Rhemrev J, Jeyendran RS, Vermeiden JP, Zaneveld LJ (1989): Human sperm selection by glass wool filtration and two-layer, discontinuous Percoll gradient centrifugation. Fertil Steril 51:685-690.
  12. Rodriguez-Martinez H (2009): In vitro fertilizing capacity of frozen-thawed bull spermatozoa selected by single-layer (glycidoxypropyltrimethoxysilane) silane-coated silica colloidal centrifugation. Reprod Domest Anim 44:390-404. https://doi.org/10.1111/j.1439-0531.2008.01081.x
  13. Rodriguez-Martinez H, Larrson B, Pertoft H (1997): Evaluation of sperm damage and techniques for sperm clean-up. Reprod Fertil 9:297-308. https://doi.org/10.1071/R96081
  14. Sharara FI, Illions EH, Coddington CC 3rd, Scott RT Jr (1995): Evaluation of the Tru-Trax cervical mucus penetration test in predicting fertilization and pregnancy rates in in-vitro fertilization. Hum Reprod 10:1481-1505.
  15. Somfai T, Bodo S, Nagy S, Papp AB, lvancsics J, Baranyai B, Gocza E, Kovacs A (2002): Effect of swim up and Percoll treatment on viability and acrosome integrity of frozen-thawed bull spermatozoa. Reprod Dom Anim 37:285-290. https://doi.org/10.1046/j.1439-0531.2002.00350.x
  16. Thundathil J, Gil J, Januskauskas A, Larsson B, Soderquist L, Mapletoft R, Rodriguez-Martinez H (1999): Relationship between the proportion of capacitated spermatozoa present in frozen-thawed bull semen and fertility with artificial insemination. Int J Androl 22:366-373. https://doi.org/10.1046/j.1365-2605.1999.00194.x
  17. Valcarcel A, de las Heras M, Moses DF, Perez LJ, Baldassarre H (1996): Comparison between Sephadex G-10 and Percoll for preparation of normospermic, asthenospermic and frozen/thawed ram semen. Anim Rep Sci 41:215-224. https://doi.org/10.1016/0378-4320(95)01453-5
  18. Watson PF (2000): The causes of reduced fertility with cryopreserved semen. Anim Report Sci 60-61:481-492.
  19. 방명걸, 정구민, 신창재, 김정구, 문신용, 장윤석, 이진용 (1993): 인간정자에 있어서 정자처리법의 비교. 대한불임학회지 20:107-115.