• Korean Journal of Animal Reproduction
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• v.21 no.4
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• pp.397-403
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• 1997

For the ultimate goal of efficient retrovirus vector-mediated transgenic animal production, we tried to increase virus titer by employing three methods: boosting virus production by treating virus-producing cells with sodium butyrate, concentration of virus stock by either filtration or ultracentrifugation. Compared to the control, applications of sodium butyrate (5 mM) treatment and filtration resulted in only 3 and 3. 6 folds of titer increases on bovine EBTr target cells, respectively. However, concentration of virus-containing medium by ultracentrifugation showed 12.5 folds of titer increase compared to the control (10${\times}$10$^5$ LacZ$^+$ TU Im), indicating the best method which can enhance retrovirus vector-mediated transgenic animal production.

• Development and Reproduction
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• v.15 no.4
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• pp.325-329
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• 2011

The main objective of this study is to investigate the distribution of coat color among Ulleung Korean Native Brindle Cattle, and to identify basic genetic elements required for the fixation of coat color traits. 1. The distribution of coat color among Ulleung Korean Native Brindle Cattle was 24.3% yellow (67/276),13.0% black (36/276), 62.7% brindle (173/276). The frequency of Brindle coat color was slightly higher (66%, 105/159) among embryo transfer from outside than that of cows produced by embryo transfer from jurisdiction. 2. Investigation of coat color distribution in each sex group showed that the frequency of yellow coat color was lower in males (18.1%, 25/138) than in females (31.4%, 43/137). On the other hand, the frequency of brindle coat color was slightly higher among males (68.1%, 94/138) than in females (56.9%, 78/137). 3. Examination of Brindle coat color expression based on the intensity of black dots on the nose indicated that the expression of black coat color increase in proportion to the intensity of black dots.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.106-106
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• 2002

미토콘드리아는 세포내의 에너지대사에서 중요한 역할을 수행하는 세포내 소기관이며, 자체의 유전물질이 모계를 통해 유전되는 특징을 가지고 있다. 포유류 초기 배아의 발생과정에서 미토콘드리아의 역할과 기능에 대한 연구는 미진한 상태이다. 본 연구에서는 생쥐 초기 배아의 발생과정에서 관찰할 수 있는 미토콘드리아 미세구조의 변화 양상을 살펴보고, 이와 초기 배아의 발생 능력과의 관련성을 밝혀보고자 하였다. 과배란 유도된 ICR 생쥐로부터 배란된 난자와 2-세포기 배아를 수획하여 76 배양액으로 포배기까지 체외배양하면서, 각각의 발생단계에 따라 시료를 수획하였다. 미토콘드리아 미세구조의 변화는 일반적인 투과전자현미경방법(TEM)을 이용하여 관찰하였다. 미토콘드리아의 미세구조는 배란 난자에서 4-세포기 배아까지는 구형이고 크리스타가 발달하지 않은 원시형태였지만, 포배기로 발달함에 따라 크리스타가 발달된 막대형의 전형적인 미토콘드리아로 분화됨이 관찰되었다. 체외배양 중에 발생이 지연되거나 정지된 배아에서 관찰한 미토콘드리아의 미세구조는 공포화 (vacuolization), 크리스타 발달 지연, 손상된 미토콘드리아의 세포막 등과 같은 비정상적인 변형을 관찰할 수 있었다. 또한, 극체에 존재하는 미토콘드리아의 미세구조는 정상적인 핵내의 유전자와의 상호작용이 없어 미분화 상태로 포배기까지 유지되는 것을 관찰하였다. 이상의 결과를 통해 미토콘드리아의 정상적인 분화 과정이 초기 배아의 발생능력과 관련되어 있음을 알 수 있었으며, 포유류 초기 배아의 체외배양시스템을 개선하는데 미토콘드리아 미세구조의 관찰과 변화에 대한 고려가 있어야 될 것으로 생각된다. buffer A 용액으로 세척하여 유리 petri dish의 바닥에 부착된 macrophage만을 cell scraper로 분리하였다. 분리한 macrophage는 0.5-1 $\times$ $10^{6}$ cells/$m\ell$가 되게 조정하여, IL-I 을 0.001, 0.01, 0.1 또한 1 ng/$m\ell$를 첨가하여 농도에 따른 효과를 조사하였고, 각각 24, 48, 72, 96 또한 120시간을 배양하여 시간에 의한 효과도 실시하였다. 각 배치구에서 얻어진 배양액은 TGF-$\beta$를 조사하기 전까지 -2$0^{\circ}C$에 동결 보존하였다. TGF-$\beta$의 측정은 TGF-$\beta$ kit(promega, USA)를 이용하여 실시하였으며, 통계학적 분석은 Anova test를 Statview program을 이용하여 분석하였다. 시험의 결과 대조구에 비해 IL-I 첨가구는 2-3배의 TGF-$\beta$생산을 보였으며, 배양시간에 따른 생산은 시간이 지남에 따라 약간 상승하는 경향을 보였으나, 유의적인 차이를 보이지는 않았다. 또한 IL-I의 농도에 따른 생산의 변화는 IL-I의 농도에 따라 약간의 차이를 보였고 유의적인 차이는 인정되지 않았다. 임신 및 비임신의 경우 임신우의 비장 macrophage가 비임신보다는 약간 상승하는 거스로 나타났다. 이상의 결과로 볼 때 IL-I $\alpha$ 는$\bet

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.183-190
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• 2006

These studies were conducted to monitor developmental competence of follicular oocytes collected from the carcass of the high meat quality in Korean native cattle using each individual protocol of IVM, IVF and IVC. The follicular oocytes that were collected from the ovaries of the cow yielded 1, $1^+\;and\;1^{++}$ meat quality were matured, fertilized and cultured using each individual protocol of IVM, IVF and IVC. As results, the number of follicular oocytes collected from individual fundamentally-registered cows yielded 1, $1^+\;and\;1^{++}$ meat grade were 28.9, 28.8 and 29.6 per head, respectively. The rates of blastocyst formation after IVM, IVF and IVC were 27.2, 28.7 and 32.9% in the cows yielded 1, $1^+\;and\;1^{++}$ meat quality, respectively. The rate of blastocyst formation was 8.4 per head. The number of follicular oocytes collected from pedigree registered cows yielded 1, $1^+\;and\;1^{++}$ meat quality were 25.8, 27.1 and 27.0 per head, respectively. The rates of blastocyst formation were 23.0, 33.7 and 42.6% in the meat quality of 1, $1^+\;and\;1^{++}$ after in vitro-manipulation, respectively (p<0.05). The rate of blastocyst formation was 8.5 per head. In conclusion, these results suggest that in vitro embryo production system using individual culture system including IVM, IVF and IVC can make good use of the gene from the carcass of the high meat quality in Korean native cattle.

• Journal of Aquaculture
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• v.6 no.1
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• pp.55-62
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• 1993

A tetraploid cyprinid loach, Misgurnus anguillicaudatus was induced by applying heat shock. The efficiency of inductions changed with the time of initiation of treatment and the duration of treatment. The highest incidence of tetraploid was obtained at the treatment of $41^{\circ}C$ for 3 min, 28 min after fertilization. Ploidy level was estimated by kariological examination and flow cytometric analysis. The tetraploid cyprinid loach had 4n = 100 chromosomes, while the diploid had 2n = 50. The erythrocytic volume of the tetraploid was 1.65 times larger than that of the diploid. The DNA content of the tetraploid and diploid showed 6.547 pg/cell and 3.067 pg/cell, respectively.

• Korean Journal of Veterinary Research
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• v.38 no.1
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• pp.209-216
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• 1998

The purpose of present study is to improve the efficiency of fusion and the developmental rates of nuclear transplanted embryos to produce genetically identical twins from Korean native cattle. The diameter of aspirated follicles had no significant effect on the recovery rates of oocytes collected by ovum pick-up technique. The fusion rates of nuclear transplanted embryos were significantly higher in 50 and $100{\mu}s$ DC duration groups(73.3 and 72.0% ; respectively) than that in $30{\mu}s$ group(55.6% ; p<0.05). The cleavage rates of nuclear transplanted embryos appeared to be significantly higher in donor nuclei derived from in vivo (65.0%) than in those from in vitro (50.5% ; p<0.01), but the developmental rates to morulae and blastocysts were not significantly different between them(13.7 vs 10.9%, respectively).

• Clinical and Experimental Reproductive Medicine
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• v.22 no.2
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• pp.105-108
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• 1995

본 연구는 혈청 무첨가 단순배양액인 CR1이 체외에서 생산된 소 수정란의 체외 배발생에 미치는 영향을 검토하고자 실시하였다. 본 연구에서 얻어진 결과를 요약해보면, 1) 총 1250개의 체외성숙 난자로 부터 체외 수정결과 본 실험의 목적상 이용될 수 있는 1,025개 (82.0%)의 분할란 (>1세포기)을 얻을 수 있었으며, 체외배양 결과 배반포기와 부화율은 각각 27.1%와 20.2%였다. 2) CR1 배양액은 소난포란 (>1세포기)의 체외발생시 난관상피세포, 난구세포, 영양배엽세포 등의 체세포와 공동배양을 유도하지 않고서도 높은 배발생율을 얻을 수 있었으며, 이러한 결과로 미루어 볼때 CR1은 난자의 체외배양시 난자성장촉진 인자를 연구하는데 효과적으로 사용될 수 있다는 것을 시사한다.

• Korean Journal of Animal Reproduction
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• v.24 no.2
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• pp.199-208
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• 2000

This study was carried out to investigate of factors concerning to ultrasound-guided follicular aspiration; level of vacuum pressure, diameter of use needle, effect of FSH hormone and conception rate after embryos transfer. 1. Oocytes collection number were 4.2$\pm$2.9 e.a to luteuml phase and follicular phase were 4.4$\pm$3.5 e.a to ovaries of Hanwoo. 2. We taked proper level of aspiration vacuum pressure was 40~120 mmHg to oocytes collection. Oocytes collected number were 4.2$\pm$3.2, 4.3$\pm$3.4, 4.5$\pm$3.4 e.a. to 40, 80, 120mmHg, respectively, follicles aspiration rate were 49, 47, 45%. 3. Effect of collection needle diameter was not difference significantly(P＜0.05), oocytes collected number were 4.4$\pm$3.5 e.a to 170 and 3.0$\pm$1.8 e.a to 18G needle, collected oocytes quality were no difference significantly (P＜0.05). 4. Follicles increase number to FSH hormone injection were 6.2$\pm$2.3 e.a to intramuscle and 1.1$\pm$2.7 e.a. to epithelial injection method. 5. Conception rate derived from E.T. was 11.1% to freezing embryos and 46.2% to fresh E.T., difference significantly(P＜0.05).

• Journal of Embryo Transfer
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• v.21 no.4
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• pp.281-286
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• 2006

This study was conducted to investigate the hormonal changes in cultured medium during in vitro culture of bovine oviduct epithelial cells (BOEC) supplemented with interleukin (IL)-4 of 0.001, 0.01, 0.1 or 1 ng/ml. BOEC were collected from the oviduct and washed 3 times with 1% antibiotic-mycotic-DMEM medium and cultured at $39^{\circ}C$, 5% $CO_2$, 95% air for 24$\sim$120 hrs. The cultured media were analyzed hormonal changes with hormonal analyzing kit (progesterone (P4), estradiol (E2) : Perkin Elmer, USA) and Transforming growth factor (TGF)-$\beta$ with Eliza kit (Promega, USA). The production of P4 in 0.001 IL-4 was increased as the culture time increased. P4 production was significantly higher in the medium cultured for 120 hrs than 24 hrs (P<0.05). P4 production in 0.01 ng/ml group was similar to that of 0.001 ng/ml. The production of E2 in 0.001 and 0.01 ng/ml groups were increased to 72 hrs like P4 production and showed significantly different between the culture periods (P<0.05). After the culture for 96 hrs, P4 and E2 production were increased to 96 hrs, but decreased at 120 hrs. The production of TGF-$\beta$ showed no changes according to culture period or supplementation of IL-4. In conclusion, the supplementation of IL-4 can increase the production of P4 and E2 and might have important role for the successful pregnancy in bovine.

• Reproductive and Developmental Biology
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• v.28 no.1
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• pp.21-27
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• 2004

This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS ＋ 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P＜0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P＜0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P＜0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P＜0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).