• Reproductive and Developmental Biology
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• 제32권2호
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• pp.117-121
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• 2008

Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma $17{\beta}$-estradiol ($E_2$) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and $E_2$ level were measured by Hemavet 950 and $E_2$ ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma $E_2$ level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma $E_2$ level, thereby causing reproductive disorders.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.250-250
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• 2004

This study was conducted to investigate the effect of the number of spermatozoa and insemination section(field) of reproductive organs at artificial insemination using laparoscopy(Fiber optic laparoscopic system, Good-Gene Co., Korea) in deer(Elk) and cattle. Twenty six elk does and fifteen cows were inserted CIDR into virginia during 12∼14 days for synchronization of estrus. (omitted)

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.73-73
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• 2001

This study was to examine whether the vitrified, one-step diluted and direct transferred Hanwoo IVM/IVF/IVC blastocysts can be successfully survived in vivo and they were succeeded into the live birth. For vitrification, blastocysts were serially exposed in glycerol (G) or/and ethylene glycol (EG) mixtures [10% (v/v) G for 5 min, 10% G plus 20% EG (v/v) for 5 min, and 25% G plus 25% EG (v/v) for 30 sect] which is diluted in 10% FBS added D-PBS. Thawing of straw was carried out in air for 10 sec and then in water bath of $25^{\circ}C$ for 20 sec. One-step dilution within the straw was done in water bath of $25^{\circ}C$ for 1 min. Vitrified and one-step diluted embryos were directly transferred into 36 (natural or hormone induced synchronized) recipient cows in 6 areas of Kyungsang Buk-Do. Pregnancies were confirmed at first when recipient cows did not return to the subsequent estrus cycle, and later by manual palpation per rectum on day 45, 90 and then living calves were derived into parturition. Overall pregnancy was 33.3%(12/36), However, higher pregnancy was obtained when the recipients exhibited estrus one day earlier than the age of transferred embryos (53.3 vs 25.0-27.3%), irrespective of synchronization methods. Also, parous recipients became pregnant higher than nulliparous heifers, And, there were not different in pregnancy rates by the aspect of corpus luteum (CL) quality of recipients (good, 29.4; fair, 37.5; poor, 33.3%). One hundred eight of frozen-thawed Hanwoo blastocysts were directly transferred into 36 recipient cows. In 12 of pregnant cows, 3 cows were aborted and 9 cows were calved [single, 66.7% (6/9): twin, 33.3% (3/9)]. Total embryo implantation rate was 11.1% (12/108). However, 9 Hanwoo calves were lived. Therefore, these results demonstrate that direct transfer technique of vitrified and one-step diluted bovine blastocysts can be applied easily and effectively with the higher pregnancy rate on field trial without the equipment and embryological skills.

• 한국수정란이식학회지
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• 제32권2호
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• pp.59-64
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• 2017

Embryo transfer (ET) could be a relevant tool for genetic improvement programs in horses similar to those already underway in other species and produce multiple foals from the same mare in one breeding season. However, there have been no reports describing equine embryo transfer performed in Korea. In the present study, we performed an equine embryo collection and transfer procedure for the first time. We examined the embryo collection and pregnancy, size of embryo during the incubation period after collection, and progesterone (P4) and estradiol-$17{\beta}$ (E2) concentrations in mare's serum at embryo collection and transfer. A total of 16 donors responded to estrus synchronization; estrus was induced in 12 donors and 4 recipients, and artificial insemination was successful in 10 donors and six blastocysts were collected from donors. Of these blastocysts, we monitored the size of blastocysts for 3 day during incubation and transferred 2 blastocysts to a recipient, with 1 successful pregnancy and foal achieved. The dimensions of equine embryo at day 7 to day 9 were $409{\mu}m$, $814{\mu}m$ and $1,200{\mu}m$. The serum P4 and E2 concentrations were $7.91{\pm}0.37ng/{\mu}L$ and $45.45{\pm}12.65ng/{\mu}L$ in the donor mare, and 1$6.06{\pm}3.27ng/{\mu}L$ and $49.13{\pm}10.09ng/{\mu}L$ in the recipient mare.

• 한국수정란이식학회지
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• 제22권2호
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• pp.89-95
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• 2007

본 연구는 재래 산양의 체세포 핵이식에 의하여 생산한 복제 산양(진순이)의 조직으로부터 공여 핵을 배양하여 다시 핵이식을 실시하여 재복제에 따른 융합율과 분할율, 이식 후의 수태율 등을 조사하여 재복제 가능성 여부를 검토하기 위하여 실시하였다. 공여 세포는 귀 유래 섬유아세포를 분리 배양하여 사용하였으며, 체내 성숙 난자는 성숙한 미경산 재래 산양에 과배란을 유기하여 외과적인 방법으로 난관 관류를 통해 회수하여 핵이식을 실시하였다. 핵이식란의 융합은 전기 자극 방법으로 실시되었으며, 융합이 완료된 핵이식란의 활성화 처리는 핵이식 3시간 후에 Ionomycin과 6-DMAP를 병용 처리하여 실시하였다. 복제 수정란의 체외 배양은 0.8% BSA가 첨가된 mSOF 배양액으로 $2{\sim}4$ 세포기까지 체외 배양을 실시한 다음 수란 산양의 난관에 외과적으로 이식하였다. 임신 진단은 발정일로 부터 제 30일과 60일째에 초음파 임신 진단기로 임신 진단을 실시하고, Progesterone농도는 이식 후 21일째와 63일째의 혈액을 채취하여 RIA 방법으로 검사하였다. 체세포 핵이식에 의한 재복제란(2nd)을 전기 자극에 의한 융합을 1회 실시하였을 때 융합율은 65.9%로서 복재란(1st)의융합을 51.0%보다 유의적(p<0.05)으로 높았으며, 2회 전기자극을 실시하였을 때는 각각 77.4 및 63.9%로서 차이가 없었으나, 3회 재복제란 융합율도 87.5%로서 복제란의 70.1%와 유의적인 차이는 없었다. 재복제 융합란의 분할율은 56.0%로j 복제 융합란의 77.7%보다 낮았다. 재복제란을 수란 산양에 이식을 실시하여 임신 제21일과 63일째 임신 진단을 실시하였을 때 수태율을 수란 산양의 발정유기 방법에 따른 수태율에 있어서 재복제란의 21일째 수태율은 39.3%로서 복제란의 17.4%보다 높았으며, 63일째는 각각 14.3 및 13.0%로서 복제 회수에 따른 수태율의 차이는 없었다. 수란 산양의 발정 유기 방법에 있어서 제 21일째에 자연발정이 발현된 수란 산양의 수태율은 45.4%로서 인위적으로 발정 동기화를 유도한 수란 산양의 35.3%보다 높았다. 제 63일째는 각각 18.2 및 11.8%로서 차이가 없었다. 이상의 결과로 볼 때 재래 산양의 체세포 핵이식에 의한 복제효율에 있어서는 복재와 재복제간에 차이가 없었으며, 수란산양의 발정 동기화 방법에 따른 수태율에 있어서도 차이가 없었다. 그러나 앞으로 재래 산양의 복제 효율 개선을 위해서는 양질 난자의 다량 확보, 산양 수정란의 체외 배양 체계 확립, 이식 기법의 개발 등에 관한 후속 연구가 이루어져야 할 것으로 생각된다.

• 한국가축번식학회지
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• 제26권1호
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• pp.69-72
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• 2002

PGF$_2$$\alpha$의 투여부위가 젖소의 발정동기화율 및 그에 따른 수태율에 미치는 영향을 구명하기 위하여 하와이주의 대규모 농장에서 사육중인 미경산 젖소에 대하여 25mg Dinoprost trometamine을 투여한 결과 다음과 같은 결과를 얻었다. 1. 둔부(근육주사)-둔부(근육주사) 처리에 있어서 첫 번째 처리된 35두 중 18두 (51.4%)가 발정이 발현되었고 그중 9두가 수태되어 60.0%의 수태율을 나타내었다. 첫 번째 처리후 발정을 나타내지 않은 개체들에 대하여 10일후에 다시 둔부에 PGF$_2$$\alpha$를 처리한 결과 14두중 8두(57.1%)가 발정이 유기 되었고 그중 4두가 수태 되어 50.0%의 수태율을 나타내었다. 2. 둔부(근육주사)-어깨부위(피하주사) 처리에 있어서 첫 번째 처리된 39두중 19두 (48.7%)가 발정이 발현되었고 그중 12두가 수태되어 70.6%의 수태율을 나타내었다. 첫 번째 처리후 발정을 나타내지 않은 개체들에 대하여 10일후에 에께부위에 PGF$_2$$\alpha$를 처리한 결과 20두중 12두(60.0%)가 발정이 유기 되 었고 그중 3두가 수태되어 25.0%의 수태율을 나타내었다. 3. 둔부 처리의 경우 점액이 유출된 27두중 18두(66.7%)가 수태로 이어졌고 점액이 유출되지 않은 13두중 5두에서 수태로 이어졌다 어깨처리의 경우에는 점액이 유출된 2두중 1두(50.0%)가 수태로 이어졌고 점액이 유출되지 않은 10두중 1두(10.0%)만이 수태로 이어졌다.

• 한국가축번식학회지
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• 제7권2호
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• pp.41-52
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• 1983

Animal in dustry in Korea urgently needs the domestic introduction and the industrial a, pp.ication of embryo transfer technique. Namely, this technique can be utilized effectively, as means of the improvement of livestocks, as means of the increase of meat production, as means of substitute for the livestock import, and dissemination of new breed. However, as this technique avaliable in our country is remaining initial stage, we can not make use of the technique industrially unless we make much improvement as follows; induction of superovulation, non-surgical recovery of embryos, synchronization between the estrus such cycles of donor and recipient, non-surgical transfer of embryos, etc. Simultaneously, the basic studies such as harvesting oocytes from ovary, in vitro culture of oocytes, in vitro capacitation of spermatozoa, cloning by culture of blastomeres and transfer of nuclei, sexing embryo, etc. should not be neglected in order to make the technique of embryo transfer more simple and convenient. For the success of these studies, universities, national and public institutes, large scale cattle farms, and small scale cattle farms should cooperate each other. For instance, universities undertake basic researches, and the national and public institutes a, pp.y the results of the researches to animal industry along with cooperation by large scale cattle farms. By the help of the cooperative organizations, the technique relevant to our environment and farm condition may be able to be finalized, and to be a, pp.ied to samll scale cattle farm. Consequently, being served to stimulate animal productivity, this technique can be contributed to the development of livestock industry in Korea.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2000년도 춘계심포지움
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• pp.64-75
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• 2000

During the last three decades considerable advances has been made in goat embryo production and transfer technology. The Korean native black goat is the most useful domestic ruminant in this country for biological investigation and application because it has a lot of merits such as relatively short generation period (1 vs 2 year for a cow), easy of handling, well adaptation, high fertility, convenient and inexpensive. This article covers the methods of superovulation, estrus synchronization, embryo collection and transfer techniques, pregnancy diagnosis and subsequent pregnancy and kidding rates for the production of transgenic Korean native black goats. More than one hundred goat kids have been produced as a result of our transgenic goat project via microinjection of foreign gene into pronuclei, in vitro culture, transfer of various stages of fresh and frozen-thawed microinjected embryos into oviducts or uteri of recipient does. We have got two transgenic goats carrying a transgene targeting the expression of recombinant human granulocyte colony stimulating factor (hG-CSF) to the mammary gland so far. Since collection and transfer of embryos in this species is usually accomplished by laparotomy, exteriorization of the reproductive tract for surgical embryo collection leads to the formation of post-operative adhesions. Nonsurgical or laparoscopic technique to reduce adhesions from repeated surgeries has great advantages in improving embryo production and transfer especially from valuable donors. We will discuss this later.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2000년도 수정란의 위생적 처리·검사 및 특수가축의 수정란이식 기술 심포지움
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• pp.64-75
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• 2000

During the last three decades considerable advances has been made in goat embryo production and transfer technology. The Korean native black goat is the most useful domestic ruminant in this country for biological investigation and application because it has a lot of merits such as relatively short generation period(1 vs 2 year for a cow), easy of handling, well adaptation, high fertility, convenient and inexpensive. This article covers the methods of superovulation, estrus synchronization, embryo collection and transfer techniques, pregnancy diagnosis and subsequent pregnancy and kidding rates for the production of transgenic Korean native black goats. More than one hundred goat kids have been produced as a result of our transgenic goat project via microinjection of foreign gene into pronuclei, in vitro culture, transfer of various stages of fresh and frozen-thawed microinjected embryos into oviducts or uteri of recipient does. We have got two transgenic goats carrying a transgene targeting the expression of recombinant human granulocyte colony stimulating factor(hG-CSF) to the mammary gland so far. Since collection and transfer of embryos in this species is usually accomplished by laparotomy, exteriorization of the reproductive tract for surgical embryo collection leads to the formation of post-operative adhesions. Nonsurgical or laparoscopic technique to reduce adhesions from repeated surgeries has great advantages in improving embryo production and transfer especially from valuable donors. We will discuss this later.

• Journal of Animal Science and Technology
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• 제66권1호
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• pp.156-166
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• 2024

Pigs are genetically, anatomically, and physiologically similar to humans. Recently, pigs are in the spotlight as a suitable source animal for xenotransplantation. However, to use pigs as source animals, pigs should be raised in designated pathogen-free facilities. There is abundant data from embryo transfer (ET) experiments using farm pigs as surrogates, but data on ET experiments using minipigs are scarce. Eighty minipigs were used for ET experiments and after transplantation, the implantation and delivery rates were investigated. It was also confirmed whether the pregnancy rate could be increased by changing the condition or surgical method of the surrogate. In the case of minipigs that gave birth, the size of the fetal sac on the 28th day of ET was also measured. The factors that can affect the pregnancy rate such as estrus synchronization program, ovulation status at the time of ET, the number of repeated ET surgeries, and the ET sites, were changed, and the differences on the pregnancy rate were observed. However there were no significant differences in pregnancy rate in minipigs. The diameter of the implanted fetal sac on the 28th day after ET in the minipigs whose delivery was confirmed was calculated to be 4.7 ± 0.5 cm. In conclusion, there were no significant differences in pregnancy rate of minipigs in the comparative experiment on various factors affecting the pregnancy rate. However, additional experiments and analyses are needed due to the large individual differences of the minipigs.