• 한국가축번식학회지
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• 제20권2호
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• pp.179-190
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• 1996

Sexing and developing from splitted embryos which were fertilized in vitro implicate a possibility of production of the superior and sex controlled individuals. This study was carried out to investigate the production of transferable late blastocysts from in vitro fertilized embryos and to analyze sex by chromosome analysis from same embryos. In results, the ratio of cleavage and fertility of bovine follicular oocytes matured in vitro was 90% in co-cultured with granulosa cells. The competence of embryonic development from in vitro matured and fertilized bovine oocytes was 38% in co-cultured with bovine oviductal epithelial cells. To produce a lot of transferable embryos, therefore, the best conditon of culture system was co-cultured with granulosa cells for immature bovine oocytes and then co-cultured with bovine oviductal eptithelial cells for matured and fertilized oocytes. In chromosome analysis, 93% of in vitro fertilized embryos were very important aspect in chromosome preparation from bovine embryos such as duration of colcemid treatment, weakening of zona pellucida, methods of hypotonic treatment and fixation treatment.

• 한국수정란이식학회지
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• 제18권2호
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• pp.151-156
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• 2003

한우의 수정란을 보존하기 위하여 고급육계통과 다유계통의 공란우를 선발하여 과배란처리에 따른 황체의 반응, 황체수에 대한 채란된 수정란의 수 그리고 채란 수정란수에 대한 이식가능한 수정란 생산에 미치는 영향을 조사하였다. 다유계통의 공란우는 이유시 체중에 대한 육종가 상위 20%이내, 고급육 계통의 공란우는 육질에 대한 육종가 상위 20%이내의 종빈우를 선발하였다. 과배란처리 방법은 발정후 11일차에 PEG 30% FSH를 견갑부에 피하주사로 1회 투여하고 13일차에 PGF$_2$a 25mg을 투여하였으며 15일차와 16일차에 각각 2회씩 인공수정하였고 23일차에 채란하였다. 과배란처리에 따른 황체의 반응은 좌측난소가 47.1% (242/513), 우측난소가 52.8%(271/513)로써 우측의 반응이 좋게 나타났고 황체의 반응과 비교하여 채란된 수정란의 회수율은 79.9%(410/513)로 나타났다. 회수된 수정란 중에서 이식이 가능한 수정란은 86.5%(355/410)이었고 수정란의 발달단계에 있어서 상실배는 61.6%(219/355), 배반포는 38.3%(136/355)로 나타났으며 두당 이식가능한 생산수는 6.4개였다.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.3-7
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• 2002

In vitro embryo culture techniques provide significant contributions not only for a basic research of fertilization and early embryogenesis, but also for a low cost mass production of bovine embryos for transfer, embryo diagnosis, nuclear cloning and the production of transgenic cows. This presentation introduces newly developed serum-free media (IVD101 and IVMD101) that are effective far high yields of transferable embryos of excellent quality from in vitro-matured and fertilized oocytes. Both serum-free media are superior to a conventional serum-containing medium on the increased rates of blastocyst formation, post-thaw embryo viability, and pregnancy after transfer. Furthermore, reduced risks of calf mortality and large calf syndrome are also observed for the serum-free-derived embryos. Serum-derived embryos contain a large number of lipid droplets and immature mitochondria in their cytoplasm that may account for the lower production of transferable embryos and poor embryo quality.

• 한국수정란이식학회지
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• 제32권4호
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• pp.319-324
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• 2017

This study is to compare the effect of estrus synchronization and embryo transfer between Korean and Mongolian cattle. Embryos were collected from 9 donors housed in Asan city in South Chungcheong Province, South Korea. Embryos were collected 9 donors from Khushaat sum, Selenge province and Bayanchandmani sum, Tov province in Mongolia. Follicle Stimulating Hormone (FSH), Controlled Internal Drug Release (CIDR) and Prostaglandin (PG) were used for superovulation. Subsequently, Artificial Insemination (AI) was done for donor cow and embryo was collected after 7 and 8 days. Collected embryos were compared between Mongolian and Korean cattle. Finally, good quality and fresh embryos were transferred to 50 and 22 recipients of cows in Korea and Mongolia respectively. The findings show that Korean native cattle each donor cow produced on an average 16.9 embryos and, 10.9 embryos were found transferable. But in case of Mongolia the average production of embryos per donor cow was 8.6 embryos and, 6.2 embryos were found transferable. Embryo collection after 7 and 8 days was not difference in embryo production in Korea. But, in Mongolia embryo production after 8 days was found more efficient than the 7 days. Korean native recipient's cows (74.6%) and Mongolian recipient's cows (71.0%) respectively were found transferable ovarian stage. The result suggested that efficiency of embryo production from the superovulation method treated of Korean cow were higher than the Mongolian cow. The pregnancy rate of Korea native cattle was 72%, which was about 10% higher than that of Mongolia cattle.

• 한국수정란이식학회지
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• 제27권4호
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• pp.237-243
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• 2012

This research was investigated the relationship between the number of the transferable embryos and estrus expression rate, BCS (Body Condition Score), which affect the nutritional state of the cow, in Holstein donor cows. CIDRs were inserted into the vaginas of twenty two head of Holstein cows, regardless of estrous cycle. Superovulation was induced using folliclar stimulating hormone (FSH). For artificial insemination, donor cows were injected with $PGF_{2{\alpha}}$ and estrus was checked about 48 hours after the injection. Then they were treated with 4 straws of semen 3 times, with 12-hour intervals. Embryos were collected by a non-surgical method 7 days after the first artificial insemination. When BCS was $$\leq_-$$2.5, the total number of collected ova was 7.3 + 1.9, which is significantly lower (p<0.05) than the numbers 15.4 + 2.8 and 15.4 + 2.1 that were obtained when BCSs were 2.75 and $$\geq_-$$3.0, respectively. Whereas the numbers of transferable embryos were 5.2 + 1.4 when BCS was $$\leq_-$$2.5, which was smaller than the numbers 6.0 + 2.1 and 8.5 + 1.8 that were obtained when BCSs were 2.75 and $$\geq_-$$3.0, respectively; however, the differences were not significant. As for estrus induction rate, the cow groups whose BCSs were 2.75 and $$\geq_-$$3.0 showed 100.0% and 95.0%, respectively. Whereas the cow group whose BCS was $$\leq_-$$2.5 showed 57.1%, and the differences were significant (p< 0.05). As for estrous expression rate, the cow groups whose BCSs were $$\leq_-$$2.5, 2.75 and $$\geq_-$$3.0 showed 100.0%, 100.0% and 85.7%, respectively; however, the differences were not significant. According to the result of this research, it is considered that the total number of collected ova and the number of transferable embryos will be affected by the nutritional state before and after in vivo embryo production and superovulation treatment, and that although the mechanism is not clear, poor stockbreeding management and nutritional level would cause the decrease of ovum recovery rate and the number of transferable embryos in high-producing cows. On the other hand, diverse researches on the superovulation treatment method that is suitable for high-producing Holstein donor cows would contribute to preventing ovarian cyclicity disorder, as well as to the early multiplication of cows with superior genes by increasing the utilization value of donor cows.

• 한국가축번식학회지
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• 제11권2호
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• pp.122-126
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• 1987

This study was carried out to investigate the effects of the glycerol equilibration methods and embryo grades before freezing on the survival rate after thawing in mouse embryos. The results obtained from this study were as follows: 1. The number of embryos of grade I(Excellent), II(Good) and III(Fair) before freezing in this study was 97(27.4%), 160(45.2%) and 97(27.4%), respectively. 2. The average survival rate of frozen-thawed embryos in 3 and 5 steps glycerol equilibration was 66.7% and 64.1%, and the rate of transfererable embryos after culture was 68.9% and 69.0%, respectively. 3. Out of embryo grade I and II before freezing, the transferable rate after thawing was 75.2% and 48.1%, respectively, and grade I embryos before freezing was higher transferable rate than that of grade II.

• Asian-Australasian Journal of Animal Sciences
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• 제19권3호
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• pp.341-346
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• 2006

Despite many studies, results of superovulation protocols are not consistent in farm animals. In this study, 151 Boer goats were superovulated to examine the factors affecting superovulation and embryo transfer (MOET). An optimal regime for superovulation treatment was identified as a 4-day treatment with decreasing dosages of 6-7 mg Chinese FSH or 240 mg Canadian FSH. The 4-day treatment with decreasing dosages of 6-7 mg Chinese FSH was, therefore, adopted to study effects of the age of does, season and repeated treatments on superovulation and embryo transfer. The best season for superovulation and embryo transfer and pregnancy was autumn, and the best age range was 12-35 months old. Within animals there were no significant differences in the number of ovulations and the rate of transferable embryos between the first and the second superovulation. However, these parameters declined significantly for the third superovulation. No marked effects of the number of ovulations on the proportion of transferable embryos were noted. The parturition rate of the recipients receiving single embryos was not different significantly from those receiving two embryos, and the kidding rate calculated from embryos transferred did not differ significantly between recipients receiving one and two embryos.

• Journal of Animal Science and Technology
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• 제62권4호
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• pp.449-459
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• 2020

Here, we investigated the effects of different nutrient requirements (NR) on blood metabolites, transferable embryo number after multiple superovulations with artificial insemination (AI), body condition score (BCS), and estrus cycle in Hanwoo cow. Nineteen Hanwoo cows were randomly divided into three groups (80%, 100%, and 120% NR, containing 6, 8, and 5 individuals, respectively) and fed based on the NR. In experiment 1, glucose, total cholesterol, triglyceride, blood urea nitrogen (BUN), aspartate aminotransferase (AST), alanine aminotransferase (ALT), non-esterified fatty acids (NEFA), albumin (ALB), and total protein (TP) were analyzed. In experiment 2, total number of recovered embryos and transferable embryos was examined after embryo recovery and multiple superovulations with AI. In experiment 3, body weight, BCS, and estrus cycle were examined. In experiment 1, total cholesterol was significantly different among the 80%, 100%, and 120% NR groups (126.5 ± 3.3, 152.6 ± 2.4, and 177.4 ± 1.8 mg/dL, respectively, p < 0.05). The triglyceride and BUN levels in the 120% NR group were significantly higher than those in the 80% and 100% groups (p < 0.05). The NEFA levels were significantly different among the 80%, 100%, and 120% NR groups (440.5 ± 18.2, 318.5 ± 23.1, and 195.1 ± 8.5 ЧEq/L, respectively, p < 0.05). The AST and TP levels in the 80% NR group were significantly lower than those in the 100% and 120% NR groups (p < 0.05). In experiment 2, the 120% NR group showed a higher percentage of transferable embryos than the 80% and 100% groups (p < 0.01). The mean body weight and BCS among the 80%, 100%, and 120% NR groups were significantly different (p < 0.05). The estrus cycle in the 80% NR group was delayed compared with the 100% and 120% NR groups (20.8 ± 0.2 and 21.2 ± 0.5 days, respectively). In conclusion, the blood metabolic tests proved that Hanwoo cows with 120% NR can produce a large number of transferable embryos. Thus, 120% NR is the appropriate feeding level for this type of cows as it results in the production of a large number of transferable embryos by multiple superovulations with AI.

• 한국수정란이식학회지
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• 제27권1호
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• pp.15-20
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• 2012

The objective of this study was to investigate the relationship between body weight, body condition score (BCS), blood urea nitrogen (BUN), glucose, cholesterol and number of transferable embryos for the purpose of improving reproductive performance in Hanwoo donors. Seventy five cows, at random stages of the estrous cycle, received a CIDR together with injection of 1mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment begann. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU = 1 ml) administered twice daily in constant doses over 4 days. On the 3rd administration of FSH, CIDR was withdrawn and 25 mg $PGF_2{\alpha}$ was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received $100{\mu}g$ GnRH at the time of 1st insemination. Embryos were recovered 7 days after the 1st insemination. In conclusion, cows with body weight < 400, 400~450 and > 450kg had number of transferable embryos of $4.2{\pm}1.7$, $6.1{\pm}2.7$ and $4.8{\pm}2.6$, cows with BCS <2.25, 2.25~2.75 and ${\geq}2.75$ had number of transferable embryos of $4.6{\pm}1.6$, $5.7{\pm}2.4$ and $5.1{\pm}2.7$ respectively. These data indicate that a body weight and BCS for superovulation of CIDR-treated Korean native cows does not affect the embryo yield.

• 한국수정란이식학회지
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• 제26권3호
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• pp.153-158
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• 2011

This study was performed in order to simplify the operation and minimize stress of donor and be readily available in the field with low cost and high quality embryos using the Direct Embryo Collection (DEC). Donors, at random stages of the estrous cycle, received a CIDR. 7 days later, 200 mg FSH was treated with 40, 30, 20, 10 mg FSH levels in declining doses twice daily by intramuscular injection for 4 days. On the 3rd day administration of FSH, 25 mg $PGF_2{\alpha}$ was administered and CIDR was withdrawn. After FSH injections were complete, donors were artificially inseminated twice at 12 hr intervals. The donor cattle received 250 ${\mu}g$ GnRH at time of 1 st insemination and embryos were recovered 8 days after the 1st insemination. Embryo collection from superovulated donors was performed to flushing by non-surgical methods of 3-way, 2-way and DEC (l-way). The average number of recovered embryos were 11.25${\pm}$0.63, 12.5${\pm}$0.65 and 11.75${\pm}$0.48 from operations of 3-way, 2-way and DEC methods, respectively. There were no significant differences among the embryo collection methods. Also, The average number of transferable embryos were 6.25${\pm}$0.48, 7.25${\pm}$0.48 and 7.25${\pm}$0.63 from each embryo collection procedures. The number of transferable embryos was no differences among the 3-way, 2-way and DEC methods, respectively. Meanwhile, the ratio of transferable embryos for all recovered embryos from DEC methods was higher as 61.7 % than 55.6 %, 58 % from methods of 3-way, 2-way. And the flushing solution required for recovering embryos by DEC method was significantly lower as 0.28${\pm}$0.32 1 than 1.8${\pm}$0.12 1, 1.75${\pm}$0.10 1 from 3-way, 2-way methods (p<0.05). Also, the time required for recovering embryos by DEC methods was significantly lower as 27${\pm}$2 min than 51${\pm}$3, 45${\pm}$2 min, respectively (p<0.05). In conclusion, these results suggest that DEC method for embryo collection may be effectively used for production of in vivo embryos using less flushing solution and, it might be effectively available in the field compared to conventional embryo recovery methods using 3-way or 2-way balloon catheter.