Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Cleavage of In Vitro Fertilized Oocytes and Viability of Sperm Cryopreserved with L-Cysteine in Korea native cattle

L-Cysteine을 첨가하여 동결-융해한 한우 정자의 생존성과 체외 수정 난자의 분할

  • Park, Bola (College of Veterinary Science, Kangwon National University) ;
  • Lee, Kung-Jin (College of Animal Life Science, Kangwon National University) ;
  • Lee, Sang-Hee (College of Animal Life Science, Kangwon National University) ;
  • Lee, Eunsong (College of Veterinary Science, Kangwon National University) ;
  • Cheong, Hee-Tae (College of Veterinary Science, Kangwon National University) ;
  • Yang, Boo-Keun (College of Animal Life Science, Kangwon National University) ;
  • Park, Choon-Keun (College of Animal Life Science, Kangwon National University)
  • 박보라 (강원대학교 수의과대학) ;
  • 이경진 (강원대학교 동물생명과학대학) ;
  • 이상희 (강원대학교 동물생명과학대학) ;
  • 이은송 (강원대학교 수의과대학) ;
  • 정희태 (강원대학교 수의과대학) ;
  • 양부근 (강원대학교 동물생명과학대학) ;
  • 박춘근 (강원대학교 동물생명과학대학)
  • Received : 2013.08.15
  • Accepted : 2013.09.03
  • Published : 2013.09.30
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Abstract

This study was designed to evaluate the effect of L-cysteine on sperm characteristics and oocyte cleavage in vitro in Korean native cattle. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk and/or 0, 5, 10 and 20 mM L-cysteine before cryopreservation. The viability in frozen-thawed sperm were estimated by SYBR14/PI double stain, acrosome damage with FITC-PNA, mitochondria intact with Rhodamin123 and hydrogen peroxide($H_2O_2$) level with carboxy-DCFDA by flow-cytometry. The developmental capacity was also assessed with cleavage rates in oocytes fertilized in vitro by frozen-thawed sperm. In results, the sperm viability was significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). In addition, acrosome damage was significantly decreased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). The mitochondria intact was also significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). On the other hand, the cleavage rates were significantly increased in 0 mM, 5 mM and 10 mM groups than 20 mM concentration of L-cysteine (p<0.05). The oocyte degeneration of oocytes were significantly decreased in 0 mM, 5 mM and 10 mM groups than in 20 mM L-cysteine group (P<0.05). However, there are no significantly differences among the L-cysteine treatment groups. We suggest that concentration of 10 mM L-cysteine have beneficial impact for sperm cryopreserved in Korean native cattle. This result also could be recommended for artificial insemination program if supported by an improvement in the fertility results and required further study.

Keywords

References

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