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http://dx.doi.org/10.12749/RDB.2013.37.3.97

Development of Semen Transport System for Cryopreservation and Fertility in Bull Sperm  

Lee, Sang-Hee (College of Animal Life Sciences, Kangwon National University)
Song, Eun-Ji (College of Animal Life Sciences, Kangwon National University)
Woo, Jea-Seok (National Institute of Animal Science, Hanwoo Ex.)
Lee, Seung-Hwan (National Institute of Animal Science, Hanwoo Ex.)
Kang, Hee-Seol (National Institute of Animal Science, Hanwoo Ex.)
Cheong, Hee-Tae (School of Veterinary Medicine, Kangwon National University)
Yang, Boo-Keun (College of Animal Life Sciences, Kangwon National University)
Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University)
Publication Information
Reproductive and Developmental Biology / v.37, no.3, 2013 , pp. 97-102 More about this Journal
Abstract
The objective of this study was to develop of semen transport system for cryopreservation and fertility in bull sperm. The ejaculated semen were diluted with Triladyl containing 20% egg-yolk for transportation. Diluted semen was transported by three methods that there were wrapping tissue (Tissue), sinking under $30^{\circ}C$ water (Water) and sinking between warm water and air (Air) methods. Semen was transported within 2 hours in $0.3^{\circ}C$. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk. And frozen-thawed sperm were estimated with SYBR14/PI double stain for viability, FITC-PNA/PI double stain for acrosome reaction analysis and Rhodamine123 double stain for mitochondrial intact assessment. In results, live sperm (SYBR+/PI-) in Air treatment group ($43.3{\pm}4.7%$) was significantly (p<0.05) higher than other treatment groups (Tissue: $16.3{\pm}2.7%$ and Water: $27.5{\pm}3.1%$), dying sperm (SYBR+/PI+) in Air treatment group ($55.6{\pm}4.7%$) was significantly lower than other treatment groups (Tissue: $77.6{\pm}3.2%$ and Water: $67.6{\pm}3.3%$) (p<0.05). Acrosome reaction in Air treatment group ($0.2{\pm}0.1%$) within live sperm (PI negative region) was significantly (p<0.05) lower than other treatment groups (Tissue: $0.7{\pm}0.2%$ and Water: $0.5{\pm}0.1%$), the acrosome reaction in Air treatment group ($28.6{\pm}2.8%$) within all sperm also was significantly lower than other treatment groups (Tissue: $44.2{\pm}1.8%$ and Water: $36.2{\pm}2.0%$) (p<0.05). And mitochondrial intact in Air treatment group within live ($97.1{\pm}0.4%$) and all ($61.9{\pm}3.3%$) sperm were significantly higher than other treatment groups (Tissue: $85.2{\pm}3.3%$, Water: $87.8{\pm}2.9%$ within live sperm and Tissue: $49.28{\pm}3.7%$, Water: $42.0{\pm}3.1%$ within all sperm) (p<0.05). Therefore, we suggest that transportation by sinking method between warm water and air was beneficial to improvement of fertility in frozen-thawed in bull semen.
Keywords
Bull sperm; Cryopreservation; Transport system; Flow cytometry;
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