• 한국가축번식학회지
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• 제22권2호
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• pp.171-176
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• 1998

본 연구는 체외에서 배양된 생쥐 완전탈출 배반포기배를 동결보존액 EFS35를 이용하여 초자화 동결하였을 때 체내발달의 적합성 여부를 조사하기 위해 실시하였다. 공시된 완전탈출 배반포기배($\theta$ 130$\mu\textrm{m}$)는 체내에서 생산된 전핵기 수정란을 5일 동안 체외배양하여 얻었으며, 10% ethylene glycol(EG)에 5분 노출한 후 EFS35(35% EG, 18% Ficoll, 0.3 M sucrose)에 30초 동안 노출하거나, 초자화 동결하였다. 융해 후, 재팽창이 이루어진 완전탈출 배반포기배는 가임신 3일된 대리모의 한쪽 또는 양쪽 자궁각(4~6개/자궁각)에 이식하였다. 대리모의 임신율과 착상율은 임신 15일재 외과적 해부로 판정하였다. 그 결과를 요약하면 다음과 같다. 융해 30분 후, 완전탈출 배반포기배의 체외생존율은 노출군(65.5%)과 동결군(54.5%)간에 유의한 차이는 없었다. 또한, 체내발달율을 조사하였던 바, 착상율에 있어서 동결군(41.0%)과 대조군(58.5%)간에 유의한 차는 없었지만, 정상산자율에서는 동결군(24.0%)의 결과가 대조군(58.3%)보다 유의하게 낮게 나타났다(p<0.05). 이러한 결과는 EFS35를 이용한 완전탈출 배반포기배의 초자화 동결은 정상산자율은 감소하였지만, 완전탈출 배반포기배의 이용 효율성을 넓히는데 이용될 수 있다는 것을 알 수 있었다.

• 한국수정란이식학회지
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• 제26권1호
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• pp.37-43
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• 2011

A total 5,946 cows from 24 dairy farms were carried out for the improvement of reproductive performance. Dairy cows in post-parturition 30 day were performed periodic reproductive examination to check for recovery of post-parturition ovary and uterus and for the early diagnosis of reproductive disease. The results obtained from this studies were as follow. The result of 1,126 cows with ovario-uterine disease were 579 slient heat and error of estrus detection (51.4%), 296 ovarian disease (26.3%), 248 uterine disease (22%), mummification and freematin were each 1 head (0.1%), respectively. Hormonal therapeutic effects were follicular cyst 81.5%, luteal cyst 90.7%, endometritis 86.9%, mucometra 90.1%, pyometra 60.9%, respectively. In cows, even if the 1st treatment fails, 2nd, 3rd treatment were performed. Therapeutic effect of 2nd, 3rd were reduced, but the number of cured cows were gradually increased. The cured cows after hormonal treatment were performed service repeatedly and the cumulative conception rate were increased. The cows treated with hormones at first service, the conception rate were follicular cyst 26%, luteal cyst 64.1%, endometritis 38.7%, mucometra 40%, pyometra 20.5%, respectively. The cumulative conception rates were increased by repeated service follicular cyst 57.3%, luteal cyst 84.6%, endometritis 67%, mucometra 75%, pyometra 64.1%, respectively.

• 한국수정란이식학회지
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• 제27권3호
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• pp.133-139
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• 2012

This study was carried out to develop the useful inducing method of estrus for Korean native cows. Under the condition of estrus induction by administering $PGF_{2{\alpha}}$ for the cows in which corpus luteum (CL) in ovaries was detected by ultrasonography, ovarian responses and the changes of progesterone ($P_4$) concentration against $PGF_{2{\alpha}}$ compared with conception rate were observed in cows and heifers. In inducing estrus administering $PGF_{2{\alpha}}$. to the cows which has corpus luteum in ovaries, ovarian reponses, the changes of progesterone concentration, and conception rate were identified and compared. The results attained from the studies were as follows. Significant decreases of CL in size over time after $PGF_{2{\alpha}}$ administration were detected in both cow and heifer groups (p<0.001), but not different between groups in the CL regression rate (p>0.05). In addition, the percentage changes relative to the plasma $P_4$ concentration on day 1 after $PGF_{2{\alpha}}$ treatment were decreased to below 1ng/ml. The growth rate of follicle was observed as 31% on day 1 and 42% on day 2 in cows, and 34% on day 1 and 97% on day 2 in heifers, resulting that growth of heifers are faster than that of cows (p<0.05). The conception rate after $PGF_{2{\alpha}}$ treatment were 60.5% and 64.2% in cows and heifers, respectively. It also indicated that the conception rate after estrus observation with $PGF_{2{\alpha}}$ injection was as high as 66.6% while that with timed-artificial insemination (TAI) regardless of the estrus observation was 56%, which means the pregnancy rate of artificial insemination after estrus observation was higher than that of TAI (p<0.05). In the result of all above, there were significant decreases in CL size and the plasma $P_4$ concentration by days but rapid growth in follicles, which has no differences in cows and heifers. The conception rate was commonly high after estrus observation and more than 50% under TAI.

• 한국수정란이식학회지
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• 제29권1호
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• pp.13-19
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• 2014

This study was carried out to study the survival rate of thawed Hanwoo embryos frozen by the slow-rate freezing or the cryotop vitrification method. Hanwoo cumulus-oocyte complexes were recovered from ovaries at a slaughter house, matured for 20~22 hours, fertilized with Hanwoo semen for 5~6 hours, and cultured for 7~9 days in $38.5^{\circ}C$, 5% $CO_2$ incubator. For freezing, Day 7~9 blastocysts were collected. Embryos for the slow-rate freezing were equilibrated in 1.8 M ethylene glycol (EG) with Dulbecco's phosphate-buffered saline (D-PBS). Programmable cell freezer was precooled down to $-7^{\circ}C$, and the straw was seeded during 8 minutes-holding time, and was cooled to $-35^{\circ}C$ at the cooling rate of $0.3^{\circ}C/min$, and then was plunged and stored in liquid nitrogen. Embryos for the cryotop vitrification were treated in TCM199 with 0.5 M sucrose, 16% EG, 16% dimethylsulfoxide (DMSO). Embryos were then loaded individually onto cryotop and plunged directly into liquid nitrogen. The survival rates of embryos frozen by these two freezing methods were evaluated at 12 to 24h post-thawing. The survival rates of frozen/thawed Hanwoo embryos by the cryotop vitrification method ($56.86{\pm}26.53%$) were slightly higher than those by the slow-rate freezing method ($55.07{\pm}26.43%$) with no significant difference. Using the cryotop vitrification and the slow-rate freezing of Hanwoo blastocysts on Day 7 following in-vitro fertilization (IVF) treatment, the survival rates of frozen/thawed Hanwoo embryos were $72.65{\pm}18.3%$ and $79.06{\pm}17.8%$, respectively. The survival rates by the cryotop vitrification were higher than those by the slow-rate freezing on both Day 8 and 9 with significantly higher survival rate on Day 9 (p<0.05). Using the cryotop vitrification and the slow-rate freezing of Hanwoo embryos to compare between three different blastocyst stages, the survival rates of the blastocyst stage embryos were $66.22{\pm}18.8%$ and $45.76{\pm}12.8%$, respectively with higher survival rate by the vitrification method (p<0.05). And the survival rate of expanded blastocysts was higher than those of early blastocysts and blastocysts in two freezing methods with significantly higher survival rate by the slow-rate freezing method (p<0.05).

• 한국수정란이식학회지
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• 제18권3호
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• pp.249-255
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• 2003

종빈돈의 번식효율 증진을 위해 후보종빈돈의 성성숙일령, 첫교배일령 및 등지방 두께 등이 번식능력에 영향을 미치는 요인을 조사$.$분석하여 우수한 후보종빈돈의 조기선발에 활용코자 수행한 결과는 다음과 같다. 1. 출생계절별 첫발정일령 및 첫수정일령은 봄에 출생한 후보종빈돈은 194.14일 및 222.05일, 여름에 출생한 후보종빈돈은 163.25일 및 193.00일, 가을에 출생한 후보종빈돈은 160.25일 및 199.20일, 겨울에 출생한 후보종빈돈은 159.72일 및 190.11일로 봄에 출생한 후보종빈돈의 첫발정일령이 겨울, 가을, 여름에 출생한 후보종빈돈보다 유의적으로 늦게 나타났다(P< 0.01). 2. 등지방두께가 13∼16mm인 후보종빈돈의 첫발정일은 180.32일, 17∼20mm인 후보종빈돈은 171.24일, 21∼23mm인 후보종빈돈은 162.20일로 등지방두께가 얇을수록 첫발정일령이 지연되었으나 유의적인 차이는 인정되지 않았다. 3. 등지방두께가 13∼16mm인 후보종빈돈의 첫수정일령은 211.12일, 17∼20mm인 후보종빈돈은 202.43일, 21∼23mm인 후보종빈돈은 195.43일로 등지방 두께가 얇을수록 첫수정일령이 지연되었으나 유의적인 차이는 없었다. 4. 첫발정일령이 160일령 이하일 때에 산자수는 9.64두였으며, 161∼180일렬인 경우 10.14두, 181∼200일령인 경우 9.56두, 201일령이상인 경우 9.13두로 첫발정일령이 161∼180일이었을 때가 산자수가 가장 많았으나 유의적인 차이는 없었다. 5. 첫수정일령이 180일령 이하일 때에 산자수는 9.13두였으며, 181∼200일령인 경우 9.75두, 201∼220일령인 경우 10.13두, 221일령 이상인 경우 9.45두로 첫수정일령이 201∼220일령이었을때가 산자수가 가장 많았으나 유의적인 차이는 없었다. 6. 첫수정시 등지방두께가 13∼16mm일 때에 산자수는 9.33두였으며, 17∼20mm인 경우 9.81두, 21∼20mm인 경우 10.17두로 첫수정시 등 지방두께가 21∼23mm이었을 때가 산자수가 가장 많았으나 유의적인 차이는 없었다.

• Clinical and Experimental Reproductive Medicine
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• 제43권3호
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• pp.164-168
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• 2016

Objective: Assisted reproductive technology has been associated with an increase in multiple pregnancies. The most effective strategy for reducing multiple pregnancies is single embryo transfer. Beginning in October 2015, the National Supporting Program for Infertility in South Korea has limited the number of embryos that can be transferred per in vitro fertilization (IVF) cycle depending on the patient's age. However, little is known regarding the effect of age and number of transferred embryos on the clinical outcomes of Korean patients. Thus, this study was performed to evaluate the effect of the number of transferred blastocysts on clinical outcomes. Methods: This study was carried out in the Fertility Center of CHA Gangnam Medical Center from January 2013 to December 2014. The clinical outcomes of 514 women who underwent the transfer of one or two blastocysts on day 5 after IVF and of 721 women who underwent the transfer of one or two vitrified-warmed blastocysts were analyzed retrospectively. Results: For both fresh and vitrified-warmed cycles, the clinical pregnancy rate and live birth or ongoing pregnancy rate were not significantly different between patients who underwent elective single blastocyst transfer (eSBT) and patients who underwent double blastocyst transfer (DBT), regardless of age. However, the multiple pregnancy rate was significantly lower in the eSBT group than in the DBT group. Conclusion: The clinical outcomes of eSBT and DBT were equivalent, but eSBT had a lower risk of multiple pregnancy and is, therefore, the best option.

• 한국수정란이식학회지
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• 제23권1호
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• pp.25-30
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• 2008

This study was performed to identify and analyze the specifically expressed plasma proteins during early pregnancy in both pregnant and non-pregnant Hanwoo. Blood samples were collected at 0 (the day of AI), 2, 3, 4, 7, and 11 weeks after AI from pregnant (n=3) and non-pregnant (n=4) Hanwoo, respectively. The hematological parameters were measured. After 2-dimensional electrophoresis using serum, normalized protein spots were selected for the significant expression variation deviated over two fold in its expression level between two groups. Among 17 spots selected, 15 were identified as albumin, IgG1 heavy chain constant region, haptoglobin, ferrochelatase, fibrinogen, hemopexin. 5 spots were expressed only in non-pregnant specific. The spot identification of 1105 and 6106 was decreased after 3 weeks from AI. However, 2/17 spots were still unidentified. Further studies are needed to analyze the function of the proteins associated with early pregnancy.

• Reproductive and Developmental Biology
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• 제36권1호
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• pp.87-93
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• 2012

The purpose of this experiment is to compare the pregnancy rate (PR) according to the state of the ovaries and uterus, according to the number of embryos transferred from cows and heifers and to investigate the method of artificial twin induction with Hanwoo $in$ $vitro$ fertilized (IVF) embryos by embryo transfer (ET). Looking at the PR according to the condition of the ovaries and uterus, the result was not influenced by the condition of the ovaries, but was significantly influenced by the state of the uterus. The PR according to the number of embryos transferred from cows was 36.8%, 53.0%, 50.5% for 1, 2, and 3 embryos respectively, and although there was a higher frequency of twin calves with 3 embryos than with 2, the calving rate was the highest with 2 embryos. In case of heifers, the transfer of 1 embryo showed the best pregnancy and calving rate, and although the PR was similar with 2 embryos (67.7 versus 66.4), in case of 2 embryos transferred there was high frequency of embryonic loss( 6.1%) occurred when a cow was diagnosed at 28 and 53 d after ET, total loss (21.3%; sum of fetal death, abortion and stillbirth after pregnant diagnosis at 60 day).

• Reproductive and Developmental Biology
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• 제36권2호
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• pp.95-101
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• 2012

The purpose of this experiment was to compare the pregnancy rate (PR) according to the state of the ovaries and uterus, according to the number of embryos transferred from cows and heifers and to investigate the method of artificial twin induction with Hanwoo in vitro fertilized (IVF) embryos by embryo transfer (ET). Looking at the PR according to the condition of the ovaries and uterus, the result was not influenced by the condition of the ovaries, but was significantly influenced by the state of the uterus. The PR according to the number of embryos transferred from cows was 36.8%, 53.0%, 50.5% for 1, 2, and 3 embryos, respectively and although there was a higher frequency of twin calves with 3 embryos than 2, the calving rate was the highest with 2 embryos. In case of heifers, the transfer of 1 embryo showed the best pregnancy and calving rate, and although the PR was similar with 2 embryos (67.7 versus 66.4), in case of 2 embryos transferred there was high frequency of embryonic loss (6.1%) occurred when a cow was diagnosed at 28 and 53 d after ET, total loss (21.3%); sum of fetal death, abortion and stillbirth after pregnant diagnosis at 60 day.

• 한국수정란이식학회지
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• 제16권1호
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• pp.35-40
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• 2001

Artificial insemination (AI) with frozen or cooled semen is widely used in commercial fields of cattle and pig. Little is known about characteristics of canine sperm after freezing or cooling. For both practical and commercial goal, the canine semen treated with cooling and freezing should be carried out to exam the fundamentals, including sperm motility, survivability and fertilizing capacity. The aim of this study, thus, was to identify the effects of extended exposure to 4$0^{\circ}C$ on canine semen by motility, survivability, acrosomal changes following different duration. Fifteen ejaculates collected by digital manipulation twice per week from 3 dogs (Shih-Tzu) were divided to 16 aliquots after adding Tris-egg yolk (TE) buffer formulated by our laboratory, and cooled from 37 to 4$^{\circ}C$, by ramp rate of 0.6$^{\circ}C$/min. Each sample was evaluated by their motility, survivability and the acrosomal status at 0h (control), 2h, 12h and 1 d~10 d, respectively. The motility of spermatozoa was graded to 6 levels using the modified method of Seager. The survivability of sperm was assessed using an epifluorescence microscope after Fert/Light (Mole-cular Probes Inc.) staining. To estimate the proportion of the spermatozoa of intact acrosome, 200 spermatozoa were assessed in randomly selected fields, using epifluorescence microscope after FITC/PSA (Sigma) staining. At 2 h after cooling, the motility of most spermatozoa were assessed to be grade 0 and 1. At 12 h, high number of sperm were in grade 0 to 1, however, it was significantly (P<0.05) lower than that of 2 h. From 1 d to 4 d, ~50% of sperm was assessed to grade 0 to 1. On day 7, a little sperm were in grade 0 to 1. No sperm showed motility on day 10. Sperm motility was rapidly reduced by the percent of 10% of grade 0 to 1. From 2 h to 6 h, the number of live sperm was 90% and the sperm chilled for 10 days lived>50%. Acrosomal intact of spermatozoa exposed to 4$^{\circ}C$ for 2 h was 51%, supposed the sperm of control was 100%. Our results suggest that 1) this is easy to transfer and preservation for short periods 2) AI can be used by semen chilled for 6-Day.