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KMID : 1023620110350030385
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Reproductive and Developmental Biology
2011 Volume.35 No. 3 p.385 ~ p.390
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Analysis of Frozen-Thawed Sperm Characteristic in Alpha 1,3-Galactosyltransferase(GalT) Knock-out Cloned Miniature Pig
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Woo Jae-Seok
Lee Yong-Seung
Yoo Han-Jun
Hwang Seong-Soo
Oh Keon-Bong
Cheong Hee-Tae
Yang Boo-Keun
Park Soo-Bong
Park Choon-Keun
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Abstract
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The purpose of this study was undertaken to evaluate of cryopreservation efficiency in α 1,3-galactosyltransferase knock-out(GalT KO) cloned miniature pig sperm. To compare ability of frozen-thawed sperm characteristics, three different pig strains (GalT KO) cloned miniature pig, PWG miniature pig and Duroc were used. The ejaculated semen from the three pig species was diluted with same volume extender and added to LEY solution for freezing. The diluted semen was placed in 0.5 ml straws, and freezing was initiated by exposing the straws to liquid nitrogen (LN2) vapours for 10 min before placing them into LN2 for cryopreservation. After thawing, the sperm ability were assessed for viability (SYBR-14/PI staining), abnormality (Rose Bengal staining), and acrosome status (intactness, intensity and capacitation) (chlorotetracycline, CTC staining). The viability of frozen-thawed GalT KO pig sperm had no significant difference as compared with Duroc and PWG miniature pig sperm. However, The CTC pattern of frozen-thawed GalT KO cloned miniature pig spermatozoa showed significantly lower rates in F pattern and AR pattern (p<0.05) and significantly higher rates in B pattern than Duroc and PWG miniature pig (p<0.05). The abnormality of GalT KO cloned miniature pig sperm was significantly lower as compared to Duroc and PWG miniature pig sperm (p<0.05). In conclusion, GalT KO cloned miniature pig semen can be cryopreserved successfully and used for artificial insemination reasonably.
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KEYWORD
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α 1, 3-Galactosyltransferase knock out(GalT KO), Cloned miniature pig, Cryopreservation, Sperm ability, Duroc, PWG miniature pig
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