• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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• pp.6-10
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• 2001

No. Sperm can be sexed with 90% accuracy by flow cytometry/cell sorting. No. The current speed of sexing is about 5,000 live sperm of each sex per second, remarkably fast considering that each sperm is individually sexed. No. Although fast, sperm sexing is not fast enough to use standard numbers of sperm per AI dose. No. With well managed heifers, pregnancy rates with low doses of sexed, frozen sperm are 70-80% of those with unsexed sperm with normal sperm numbers. Pregnancy rates are lower in lactating dairy cows. No. Calves from sexed sperm appear to be normal. No. Sexed, frozen semen from a few bulls currently is available commercially in the United Kingdom, and likely will be available in several other countries in 2002, probably at a premium of US ＄30-50 per straw. (omitted)

• Reproductive and Developmental Biology
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• 제29권4호
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• pp.235-239
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• 2005

This study was conducted to test whether the transgenic cattle pass the transgene to their progeny through germ cells, and whether the transgene is expressed in the mammary gland of ransgenic cows. Two male ransgenic calves were born from IVF-derived embryos injected with bovine $\beta-casein/human$ lactoferrin fusion gene and then grew up to be reproducible. Semen was collected from a transgenic bull after 18 mon of age and then frozen. Bovine oocytes matured in vitro were fertilized with spermatozoa of the transgenic bull and cultured in $50\;{\mu}L$ drops of CRlaa medium supplemented with 3 mg/mL BSA. After 48 h of culture, cleaved embryos were determined for the presence of transgenes by DNA polymerase chain reaction (PCR). Proportion of transgene positives among bovine embryos fertilized with sperm of the transgenic bull was $20.9\%$ (28/134). One of transgenic bulls did not produce transgenic sperm. Out of 34 calves produced from recipient heifers inseminated with semen of the other bull, 3 $(8.8\%)$ were transgenic animals (2 females and 1 male). Thus, one transgenic bull showed a low transmission frequency below Mendelian levels in both the IVF-derived embryos and his progeny. It was demonstrated by Southern blot that copy numbers of the transgene in the transgenic progeny enhanced about 1.8 times as compared to those of the founder bull The results demonstrate that the transgenic bull carrying human lactoferrin gene could pass his transgene to the progeny through germ cells, although he is a germ-line mosaic.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.14-14
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• 2002

The purpose of this study was to determine the effect of bST treatment on progesteron concentration, embryo recovery and pregnancy rate following embryo transfer. Donor cows were superovulated with Folltropin-V and PGF₂ α combination method and then inseminated with frozen semen 3 times 12 hrs interval. Donor and recipient cows were assigned to control and bST group, of which was given a single injection of bST (500 ㎎, sc) at insemination or estrus detection. Embryo collection of superovulated cows were flushed nonsurgical method at 7 to 8 days after artificial insemination. (omitted)

• 한국가축번식학회지
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• 제23권3호
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• pp.191-203
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• 1999

본 연구는 rBST, Vito E 및 Se 투여가 한우 종모우의 정액성상에 미치는 효과를 검토하였고 정액의 동결보존 시 정액희석액에 taurine의 첨가가 융해 후 정액성상에 미치는 영향을 검토하였다. 1. rBST, Vit. E, Se 및 Vit. E와 Se 혼합 투여가 한우 종모우의 정액량, 정자농도 및 수소이온농도 (pH)에 미치는 영향을 조사한 결과, 정액량과 수소이온농도는 처리구간에 커다란 차이가 없었으며(P＜0.05), 정자의 평균농도는 각각 13.50$\pm$2.32, 16.26$\pm$2.65, 17.07$\pm$2.61, 19.23$\pm$2.07 및 19.46$\pm$2.06$\times$$10^{8}$ 정자/$m\ell$로서 Se 투여구와 혼합투여구가 대조구, rBST 투여구 및 Vit. E 투여구보다 통계적으로 유의하게 높은 정자농도를 나타냈다 (P＜0.05). 2. rBST. Vit. E 및 Se 투여가 한우 종모우의 정액의 동결융해시 정액성상에 미치는 영향을 조사한 결과, 정자의 운동율과 기형율은 투여구간에 통계적 유의차는 없었으나, 정자의 생존율은 혼합투여구가 48.24%로서 대조구 (44.48%)보다 통계적으로 유의하게 높은 생존율을 나타냈으며 (P＜0.05), rBST 투여구, Vit. E 투여구 및 Se 투여구는 각각 46.48%, 47.28% 및 46.34%로서 대조구와 통계적 유의차는 인정되지 않았다. 3. 정액의 동결보존시 정액회석액에 taurine의 첨가가 정액의 융해 후 정액성상에 미치는 영향을 조사한 결과는 총정자수, 운동율, 생존율 및 기형율에서 taurine 첨가구가 taurine 무첨가구보다 다소 좋은 결과를 얻었지만 커다란 차이가 없었다. 4. rBST, Vit. E 및 Se 투여가 정자의 수정능획득과 첨체반응에 미치는 효과를 조사한 결과, 수정능 획득과 침체반응이 모두 일어나지 않는 비율(F율)과 모두 일어난 비율 (AR율) 은 투여구간에 커다란 차이가 없었다 (P＜0.05). 5. 정액의 동결보존 시 taurine 의 첨가가 융해후 정자의 수정능획득과 첨체반응에 미치는 영향을 조사한 결과는 수정능 획득이 일어났지만 첨체 반응이 일어나지 않은 비율 (B율)과 AR 율에서는 투여구간에 커다란 차이가 없었지만, F율은 taurine 첨가구가 taurine 무첨가구보다 낮은 비율을 나타났다 (P＜0.05).

• 한국임상수의학회지
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• 제7권2호
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• pp.517-519
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• 1990

소 난소로부터 난자를 채취하여 체외성숙과 체외수정을 실시하였다. 난의 성숙배양을 위한 배양액은 소 태아혈청을 첨가한 TCM 199으로 $CO_2$배양기에서 24시간 배양하였다. 체외수정에는 동결융해정액을 사용하였으며 Caffeine과 Ca Ionophore 처리후 매정에 공하였다. 배양난자중 40%가 Metaphase II까지 성숙하였다. 성숙율에 대한 소 태아혈청의 농도변화와 HEPES 첨가여부의 효과는 없었다. 체외수정율과 분할율의 성적은 각각 23.1%와 14.4%였다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권9호
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• pp.1411-1420
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• 2020

Objective: The aim of study was to investigate the effects of season and single layer centrifugation (SLC) before cryopreservation on post-thaw bull sperm quality in Thailand. Methods: Semen was collected from 6 bulls (Bos indicus) in summer, rainy season and winter 2014 through 2016. Semen characteristics, sperm morphology, sperm kinematics, viability, chromatin structure and mitochondrial membrane were evaluated. Meteorological data were available from the local meteorological station; Results: Season had an effect on semen characteristics in the raw ejaculate, with higher proportions of normal spermatozoa and lower abnormalities in winter than in the other two seasons. Sperm kinematics, viability, DNA fragmentation index, and mitochondrial membrane potential were not different between seasons. Sperm samples selected by SLC had greater normal morphology and a lower proportion with bent tails than controls and higher values of progressive motility (PRO), beat cross frequency, linearity, straightness, wobble (WOB), and lower values of slow motility, velocity average path (VAP), velocity curved line, and amplitude of lateral head displacement than controls. In addition, SLC-selection had a favorable effect on PRO, VAP, and WOB that differed among seasons. Conclusion: Our results suggested that these bulls were well adapted to their location, with season having an effect on sperm morphology. Moreover, SLC could be used prior to cryopreservation, regardless of season, to enhance normal morphology and kinematics of bull sperm samples without adversely affecting other parameters of sperm quality. However, there was considerable variation among bulls in DNA fragmentation index, mitochondrial membrane potential and sperm viability. In addition, SLC had a positive effect on sperm morphology and sperm kinematics, which could be expected to influence fertility.

• 대한수의학회지
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• 제33권1호
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• pp.179-188
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• 1993

The developmental capacity of bovine oocytes under three different culture systems was investigated in this experiment ; One was culture in TCM199 with bovine oviductal epithelial cells(BOEC) for in vitro culture, another was culture in TCM199 with BOEC for 2 days and then transfer of 4~8cell embryos to rabbit oviduct(RO) and the other was transfer of 1 or 2cell embryos to RO for in vivo culture. And the other concern of this experiment was to investigate the effect of culture period and transfer site on recovery. Immature bovine oocytes were cultured in TCM199 with granulosa cells for 22-24hrs and then fertilized in vitro using frozen-thawed semen treated with BO-caffine and BO-BSA. Fifteen to 18hrs after in vitro fertilization oocytes were cultured in TCM199 with BOEC or transferred to RO for 5 days. The rate of development to the morula or blastocyst was higher in transfer of 1 or 2cell embryos to RO(23.1%) than culture in TCM199 with BOEC(11.7%). But, there was no difference between transfer of 1 or 2cell embryos and transfer of 4~8cell embryos to RO(12.8%). Recovery under different culture periods in RO was significantly higher in 90~95hrs(70.1%) than 122~125hrs(50.9%, p<0.05) and recovery significantly increased when oocytes were transferred deeper in RO(2.5cm>, 47.7% ; 2.5~4.5cm, 63.9% ; 4.5cm<, 77.3%, p<0.05). The results show that transfer of 1 or 2cell embryos to RO is an effective means of supporting the further development of in vitro matured and fertilized bovine oocytes than culture in TCM199 with BOEC or transfer of 4~8cell embryos to RO, and recovery from RO increases when oocytes are transferred deeper and incubated shorter in RO.

• 한국수정란이식학회지
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• 제17권1호
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• pp.55-59
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• 2002

ICSI시 동결 융해한 부고환 정자의 이용 가능성을 알아보고자 난자의 배양시 체외성숙율과 활성화 처리를 한 난자와 동결 융해한 부고환 정자로 ICSI시 체외발생율을 조사하였으며, 결과를 요약하면 다음과 같다. 1. 난포란을 회수 후 24시간 배양하였을 때 배양 시간에 따른 GV, MI, M II로의 체외성숙율은 각각 7/60(11.7%), 5/60(8.3%), 48/60(80.0%)였고 30시간 배양 시간에 따른 GV, MI, M II로의 체외성숙율은 각각 3/60(5.0%), 4/60(6.7%), 53/60(88.3%)였고 퇴화란은 각각 2/60(3.3%)와 1/60(1.7%)였다. 2. 동결 융해한 부고환 정자를 이용하여 활성화 처리를 한 난자에 ICSI를 하였을 때 상실배와 배반포로의 체외발생율은 각각 12/46(26.1%), 22/46 (47.8%)로서 비활성화처리 난자군 5/39 (12.8%), 10/39(25.6%)에 비해 높은 체외발생율을 나타냈다. 3. 활성화 처리를 한 난자에 신선정자, 부고환 정자 및 동결 융해한 부고환 정자로 ICSI시 체외 발생율은 각각 24/45(53.3%), 15/40(37.50%), l1/43 (25.6%)로서 신선정자에 비해 동결 융해한 부고환 정자처리군은 체외발생율은 약간 낮았지만 이용 가능성이 있음을 확인하였다.

• 한국임상수의학회지
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• 제9권1호
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• pp.323-332
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• 1992

The present study was carried out to examine the effect of oviduct epithelium and its conditioned medium on e development of early bovine embryos in vitro. Oocytes obtained from ovarian follicles of slaughtered cows were cultured in TCM199 with 10% fetal calf serum for 22-24hrs and then fertillzed in vitro using frozen-thawed semen treated with BO-caffein, BO-BSA(20mM heparin added). Oviduct epithelium was collected in each stage of the estrus cycle and conditioned medium was the medium in which oviduct epithelium in early luteal stage was cultured. In vitro fertilized bovine embryos of 1～2 cell were co-cultured with oviduct epithelium from different estrus cycles, cultured in conditioned medium, and cultured in rabbit oviduct. The cleavage rates of in vitro fertilized early bovine embryos co-cultured with oviduct epithelial cell from early luteal, luteal and follicular phase of estrus cycle(67.2～70.8%) and cultured in conditioned medium(56.7%) were significantly(p<0.05) higher than that of the control(44.2%) The rate of development to morula or blastocyst stage in oviduct epithelial cell co-culture(15.3～32.5%) from three phase of estrus cycles and conditioned medium(14.5%) were significantly(p<0.05) higher than that of the control(5.2%). The oviduct epithelial cell from early luteal phase gave a significantly( p<0.05) higher rate of development to morula or blastocyst stage than both luteal and follicular phase. The results of in vivo culture in rabbit oviduct of early bovine embryos were 52.1% for the cleavage rate and 26.7% for the rate of development to morula or blastocyst stage.

• 한국수정란이식학회지
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• 제14권2호
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• pp.113-119
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• 1999

This study was performed to improve the development of the in vitro fertilized bovine embryos by the condition of in vitro maturation. COCs were matured in TCM 199 supplemented with 0.1% PVA, 10ng/ml EGF, Hormones (5$\mu\textrm{g}$/ml FSH, 10 IU hCG, 1 $\mu\textrm{g}$/ml estradiol 17-$\beta$) or granulsa cell+Hormones atmosphere 39$^{\circ}C$, 5% CO2, 95% air for 24hrs. Matured oocytes were fertilized with frozen-thawed semen capacitated with 5mM caffein in BO medium for 20 hrs. IVF embryos were cultured in TCM 199 containing with hormones(same as matured medium), 10% FBS and co-culture with bovine oviduct epitherial cells. Maturation rates of COCs were showed 73.8%, 78.5%, 83.2% and 87.6% respectively, and were significant differences between PVA, EGF, and Hormones, GC+Hormones(p<0.05). The cleavage rates of IVF embryos were revealed 72.5%, 78.4%, 82.3% and 84.2% and showed same tendency as maturation rates(p<0.05). The blastocysts matured by above maturation condition and cultured for 7~10 days after fertilization had 34.4, 43.6, 52.3 and 59.3 cells had no differences among the treatments. These results suggest that high molecules as a substitutes of serum and growth factor may induce nuclear resumption of COCs but we need more study to produce transferable IVF blastocysts by use of that agents.