• 대한수의학회지
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• 제34권2호
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• pp.395-406
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• 1994

To study the conditions to enhance success of embryo transfer in the dog, 20 mixed-breed bitches were used for the experiment along with 4 male dogs for mating. The bitches were paired according to synchronism of natural estrus, or the counterpart as donor or recipient was treated with gonadotropin as FSH (follicular stimulating hormone) or PMSG (pregnant mare serum gonadotropin) for induction of estrus to be synchronized with estrus of the other bitch in natural estrus. Embryo recovery was performed in two ways for comparison, either by flushing each uterine horn after ovariohysterectomy or by flushing each horn in the state of non-ovariohysterectomy. In addition, the result of pregnancy according to the embryo stage and the repeatability of the experimental animals as donor or recipient were also investigated. FSH or PMSG was administered to the bitches which had passed over 4 months from last estrus, resulting in estrus-positive in 3 dogs of 6 FSH-treated dogs (50.0%), and in 5 dogs of 9 PMSG-treated dogs (55.6%), determined by proestrus signs and vaginal smear test. Estrus-positive bitches induced with gonadotropin were used as donor or recipient resulting in one embryo-recovered bitch as donor and one offspring-delivered bitch as recipient in 5 PMSG-treated dogs, whereas no result was obtained from 3 FSH-treated dogs. The rate of embryo recovery to be compared with number of corpus luteum was 68.2% in ovariohysterectomized dogs and 55.2% in non-ovariohysterectomized dogs, respectively. The number of dogs from which embryo was collected were 4 dogs of 6 ovariohysterectomized dogs (66.7%) and 6 dogs of 7 non-ovariohysterectomized dogs (85.7%), respectively. The result of parturition was obtained from one dog of 5 estrus-induced recipients, whereas no result was obtained from 3 natural-estrus recipients. The only dog which delivered a male puppy had been transferred 3 morulae and 2 blastocysts. Of 6 repeat-used bitches in canine embryo transfer, 3 dogs showed repeatability either as donor or recipient. These results indicated that inducing estrus of a dog with gonadotropin is feasible in canine embryo transfer to be synchronized with that of a natural-estrus dog, that embryo recovery is also possible in non-hysterectomized dogs, that the estrus-induced dog is also usable as recipient to result in parturition, and that repeat-use of a bitch as donor or(and) recipient is possible in canine embryo transfer.

• 한국가축번식학회지
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• 제18권1호
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• pp.63-70
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• 1994

30마리의 암양에게 surgical embryo collection과정을 통해 oviduct와 uterine horn에 trauma를 생성시켰다. 수술시 절개부위를 봉합하기 직전 노출된 abdominal tissue에 irrigigant로 10% dexamethasone을 사용하였다. 처리군에게는 17mg의 colchicine(1ml/ewe)를 투여했으며, 대조군에게는 1.0ml의 placebo를 처리하였다. 처음 17mg/im의 colchicine를 처리한 15마리 양은 colchicine독성증세를 2∼5일 내에 보였기 때문에 본 연구에서 제외되었다. 17mg에서의 colchicine독성 때문에 colchicine수준은 8, 4 그리고 2mg으로 낮추어졌다. 8mg group에 있던 또 하나의 양은 5일째에 독성증세를 보였기 때문에 나머지 양들은 4와 2mg의 수준으로 이틀에 한번씩 처리되었다. 두 번째 laparotomy는 첫 번째 처리로부터 9주후에 실시되었다. 두 번째 laparotomy후 처리군은 4mg의 colchicine을 이틀에 한번씩 14일 동안 처리되었는데 아무런 독성증세를 나타내지 않았다. 세 번째 laparotomy는 마지막 처리 5주후에 실시되었고 adhesion score로 계산하였다. Adhesion grading은 0∼4의 분포에 근거하여, 4는 가장 심한 adhesion을 나타낸다. 두 번째 laparotomy 결과 adhesion grading( 3)은 두 group 사이에 큰 차리가 없었다(P>0.05). 세 번째 laparotomy결과는 처리군에서 약간 낮은 수치를 보였지만, 통계적으로 두 grouprks에는 큰 차이는 없었다(P>0.05). 10마리의 양(5마리는 대조구, 5마리는 처리구)들은 처리후 5일경에 bone marrow analysis를 통해 cytogenetically분석되었다. 두 grouprks 염색체수와 구조에 있어서 차이가 없었다(p>0.05).

• 대한수의학회지
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• 제36권3호
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• pp.719-729
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• 1996

To investigate changes of estrus signs and genital organs in the bitch by hormonal induction of estrus, fourteen bitches of nulliparous and multiparous(2nd-5th) were grouped into diestrus and anestrus according to their estrus cycle. The hormonal treatments were divided into four groups: group A($PGF_2{\alpha}+PMSG+hCG$) and group B(PMSG+hCG) in diestrus bitches and group C(GnRH+FSH+hCG) and group D(PMSG+hCG) in anestrus bitches. The external signs of proestrus and estrus as well as the vaginal smear findings and natural breeding as estrus detection were investigated in all the experimental groups. Also, genital organs were examined at two months after the hormone treatment. The bitches in anestrus showed 100% of male attraction, vaginal bleeding and vulvar swelling as proestrus signs after the hormonal treatment for estrus induction and they showed higher numerical value of signs than the bitches in diestrus. The group A showed the lowest value in proestrous signs of all the groups. The bitches in anestrus treated with GnRH+FSH showed 100% of positive estrus by vaginal smear findings and 75% of natural breeding as estrus detection index and these values were the highest of all the groups. Pregnancy was recognized in only group C and the conception rate was 7.14% in al the experimental animals. Of the side effects after the hormone treatment, external findings of continous male attraction, continous external swelling and purulent exudate were recognized in all the experimental groups and the bitches in diestrus showed higher value of the findings than the bitches in anestrus. Of the changes of genital organs after the hormone treatment, hypertrophy of uterine horn, sanguineous exudate and purulent exudate as uterine findings were recognized in all the groups and these findings were shown more in the bitches in diestrus than in those in anestrus. These results indicated that group C showed the highest value of all the experimental groups in external signs of estrus and estrus detection and also pregnancy was recognized only in that group, consequently, that the hormonal treatment of group C would be the most effective for estrus induction, and also indicated that bitches in anestrus were more suitable than bitches in diestrus for the induction of estrus. In addition, side effects in external genital organs and uteri after hormone treatment were shown more in the bitches in diestrus than in those in anestrus, indicating that bitches in anestrus would be of choice for estrus induction.

• 한국수정란이식학회지
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• 제23권3호
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• pp.217-222
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• 2008

In previous studies, we reported that sow which was transferred OPS-freezing embryos not able to deliver a piglet (Kim et al, 2004). This study was conducted to investigate a possibility of gilt as recipients which produce piglets after transfer of OPS-freezing embryos. All transferred embryos were prepared by in vitro production (IVP) system. In vitro culture (IVC) medium used glucose-free NCSU23 supplemented with 5mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$. From day 3 of IVC, 10% fetal bovine serum albumin was added to the culture medium. In preparing of freezing embryos, embryos were treated with 7.5 $\mu g/ml$ cytochalasin-B for 30 min and centrifuged at $13,000{\times}g$ for 13 min. And then, embryos were exposed sequentially to an ethylene glycol (EG) solution, aspirated into open pulled straw (OPS), and plunged or thawed into the liquid nitrogen. In embryo transfer (ET), we used two kinds of type (surgical method vs. non-surgical method). In surgical method of embryo transfer, $55\sim65$ embryo were transferred in both uterine horn of two recipient gilts by plastic straw. Non-surgical method which is like artificial insemination was performed on three gilts. Each 140 frozen embryos were transferred to two gilts and 40 fresh embryos to one gilt. Pregnancy establishment was shown one recipient at 45 days after ET. However, the one recipient was also aborted at 58 days after ET. These results suggest that gilts can be considered as a candidate of recipients for OPS-freezing embryo transfer.

• 한국동물생명공학회지
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• 제34권3호
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• pp.240-246
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• 2019

The aim of this study was to investigate effect of heat stress on expression levels of plasminogen activators (PAs) related mRNAs and proteins, and changes of PAs activity in porcine endometrial explants. The endometrial explants (200 ± 50 mg) were isolated from middle part of uterine horn at follicular phase (Day 19-21) and were pre-incubated in serum-free culture medium at 38.5℃ in 5% CO₂ for 18 h. Then, the tissues were transferred into fresh medium and were cultured at different temperature (38.5, 39.5, 40.5 or 41.5℃) for 24 h. The expression level of urokinase-type PA (uPA), type-1 PA inhibitor (PAI-1), type-2 PAI (PAI-2), and heat shock protein-90 (HSP-90) mRNA were analysis by reverse-transcription PCR and proteins were measured by western blotting. The supernatant were used for measurement of PAs activity. In results, mRNA and protein levels of HSP-90 was higher in 41.5℃ treatment groups than other treatment groups (p < 0.05). The expression of uPA, PAI-1, and PAI-2 mRNA were slightly increased by heat stress, however, there were no significant difference. Heat stress condition suppressed expression of active uPA and PAI-2 proteins (p < 0.05), whereas PAI-1 protein was increased (p < 0.01). Although PAI-1 protein was increased and active uPA was decreased, PAs activity was greatly enhanced by exposure of heat stress (p < 0.05). These results suggest that heat stress condition could change intrauterine microenvironment through regulation of PAs activity and other factors regarding with activation of PAs might be regulate by heat stress. Therefore, more studies regarding with regulatory mechanism of PAs activation are needed.

• 한국임상수의학회지
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• 제17권1호
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• pp.234-237
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• 2000

The dornor, 2 years old, 20kg and mixed breed, was bred naturally on day 1 and day 3 of estrus and eight gastrulae were collected by flushing the uterus of the donor after laparatomy on day 13 after the second mating. The embryos were frozen by programmable freezer and preserved for about 3 months in liquid nitrogen. Another bitch in natural estrus, 2 years old, 30kg, mixed breed, was selected as the recipient and the frozen embryos(8 gastrulae) were thawed and each 4 embryos were transferred into upper partr of left and right uterine horn, respectively, on day 13 after the proper mating day determined by vaginal smear. The ecipient delivered 6 offspring 48 days after embryo transfer. Of 6 puppies, one was still birth and two puppies died one month after birth. Parentage test was performed by DNA analysis using microsatellite sequences for 3 puppiers, the recipient, the donor, the male dog bred with the donor, and the male dog raised near to the recipient. The markers selected for the test were CXX 873(133-157 base pair) and CXX 894(141-165 base pair). Using primers manufactured according to the markers, the blood samples were processed for polymerase chain reaction and the PCR products were treated for electrophoresis. The three puppies showed identical band to that of recipient, consequently, it was concluded that the puppies were offspring of the recipient mated naturally by the male dog, not the offspring by embryo transfer.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.77-77
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• 2003

Chimeric animals are referred to as an organism composed of tissues derived from more than one species. In order to examine if a pluripotency of embryonic stem cells can cross the limitation of a species, we tried to establish human-mouse chimeric animals. Human embryonic stem cells were genetically modified to express eGFP using eukaryonic expression vector pcDNA 3.1 (In Vitrogene) for an easy identification. After selection with neomycin, approximately 15 cells were implanted into mouse blastocoele cavity. Ten chimeric blastocysts were transferred to one of the uterine horn of 2.5 days pesudopregnent ICR female. Out of 272 blastocysts transferred to pseudopregnant recipients 20 live newborn were obtained after 20 days. When newborn were obtained, pups were quickly removed immersed into 4% PFA. By histological examination using fluorescent microscope, green fluorescence was observed from the liver, heart, and spleen in newborn mice. Three weeks after born, presence of eGFP sequence within mouse genome (tail and kidney) was reconfirmed by PCR. eGFP sequence was amplified from the progenies of the animal suggesting a genetic transmission of the transgene. These chimeric mice having human cells at the beginning of development, are expected to recognize human cells as “self”, therefore, human cells or tissues will be able to escape the immunological surveillance of the host if grafted into the animal. These animals will serve as a good model system for studying the graft rejection in tissue transplantation and the potential of the cells to work well in many human disease.

• 한국수정란이식학회지
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• 제15권3호
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• pp.255-261
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• 2000

The objective of this study was to improve the efficiency of bovine embryo transfer by transferring of Hanwoo embryos into Hanwoo or Holstein recipients. The cryopreserved or fresh in vitro produced(IVP) embryos were transferred into uterine horn contralaterally or ipsilaterally to the corpus luteum. The recipients were inseminated by artificially on the next day of estrus. The pregnancy was diagnosed by rectal palpation at 60∼90 days after transfer of the embryos. The pregnancy rate by transfer of one or two embryos was 78%(7/9) and 74%(31/42), respectively. The pregnancy rates according to the grade of corpus lutea of recipients was 75% (20/27) and 82.0%(18/22) at the grade of A and B, respectively. Ten(67.0%) of 15 Holstein recipients transferred with IVP Hanwoo embryos and 5(42.0%) of 12 Holstein recipients transferred with frozen IVP Hanwoo embryos were pregnant. The single and twin calving ratio in Hanwoos was 77.0%(10/13) and 23.0%(3.13) in the recipients transferred with IVP embryos and 64.0%(7/10) and 27.0%(3/10) in the recipients transferred with frozen IVP embryos, respectively. Twenty-four pregnant cows following transfer of IVP embryos, 21(88.0%) calved the normal calves, and 2(8.3%) aborted. When the frozen IVP embryos were transferred, 16 pregnant cows calved 14(88.0%) normal calves and 2(13.0%) aborted. In conclusion, when one or two IVP bovine embryos were transferred into recipients, the A and B grade of corpus luteum resulted in high pregnancy rates. For the production of twin calves, transfer of the IVP or frozen IVP embryos could be suitable.

• 대한수의학회지
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• 제29권1호
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• pp.61-68
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• 1989

This study was carried out to determine suitable selection factors for recipients and embryos which could improve pregnancy rates following bovine embryo transfer. The experiment included 52 surgical transfers from February, 1985 through June, 1986 performed on Kyungpook Breeding Center in southern Korea. The pregnancy rate was highest when recipients were in estrus within 6 hours before the donor to 12 hours after the donor (78.3% versus 50% for recipients in estrus earlier or later). Pregnancy rates were acceptable following culture under field conditions for up to 17 hours. More recipients over 15 months of age (76.1%) remained pregnant than those under 15 months (66.7%). Embryos transferred during the months from February to July resulted in higher pregnancy rates than those transferred during the remaining 6 months (77.3% versus 57.1%). Transferrable embryos were classified A (best) to C (worst); those graded A or B resulted in significantly higher pregnancy rates than those graded C (81.8% and 73.3% versus 25.0%, p<.05). Pregnancy rates among recipients of the Korean native breed tended to be higher than among Holstein recipients (100% versus 71.1%). Similarly, when the embryo was transferred to the right uterine horn, pregnancy rates tended to be higher than when it was transferred to the left (81.3% versus 65%). Pregnancy rates did not differ according to the stage of development of the embryo; they were for morulae, tight morulae, blastocysts, and advanced blastocysts, respectively: 75.0%, 66.7%, 75.0%, and 77.4%.

• 한국수정란이식학회지
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• 제13권1호
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• pp.61-67
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• 1998

This study was carried out to establish an effective system for embryo transfer techniques by analyzing several factors affecting in-vivo embryo transfer in Korean cattle. Embryos produced in-vivo were transferred into a total of 301 recipients The results obtained in studies on the factors affacting pregnancy rate after embryo transfer by condition of recipients were as follows ; 1. The pregnancy rate by age and parity of recipients showed high in 5~8 and over 12 years old(72.7~73.9%), and 3rd~4th parity(82.1%) for fresh embryos(P<0.05). The pregnancy rate did not differ by age and parity of recipients in frozen embryos. The pregnancy rate of frozen embryos tended to be similar to that of fresh embryos(38.5% and 25.0~36.7%). 2. The number of observation for normal estrus cycles of recipients did not differ In pregnancy rate between one and 2 times in fresh embryos(64.9%, 69.8%). The pregnancy rate by transferred frozen embryos showed significantly higher after 2 times of observation(P<0.05, 16.3%, 37.5%). The pregnancy rate by days open did not differ between fresh and frozen embryos. But the pregnancy rate was slightly higher in 12 months and 6 months of days open for fresh and frozen embryos, respectively(70.1~71.1% and 24.5%, respectively). 3. The pregnancy rate of transferred fresh and frozen embryos into right and left side of uterine horn did not differ(62.1% : 65.9% 25.0% : 24.3%, respectively). The pregnancy rate by the grade of CL was not different in fresh embryos, but the pregnancy rate was significantly higher in the grade A than B for frozen embryos(P<0.01, 43.2%, 16.2%).