• 한국수정란이식학회지
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• 제8권2호
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• pp.143-149
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• 1993

The objective of this study was to produce calves derived from in vitro fertilization of in vitro matured follicular oocytes. Oocytes aspirated from small antral folicles of ovaries obtained at a local slaughter house were matured and fertilized in vitro. At l8hrs after insemination with Korean native cattle semen, oocytes were co-cultured for 6~7 days by utilizing co-culture system with bovine oviduct epithelial cell. After co-culture, good or excellent quality late morulae or early blastocysts were selected by morphological criteria under stereo microscope. Selected embryos were transferred to recipients on day 6 or 7 (estrus = day 0). Recipients were monitored by observation for estrus and rectal palpation after 60 days from embryo transfer. One of them went to term with the birth of a calf. This case is the first production of calf derived from in vitro fertilization in Korea.

• Clinical and Experimental Reproductive Medicine
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• 제40권3호
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• pp.122-125
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• 2013

Objective: The majority of embryo transfers (ETs) to date have been performed on day 3 to reduce the potential risk of developmental arrest of in vitro cultured embryos before ET. Development of sequential media has significantly improved culture conditions and allowed blastocyst transfer on day 5. While day 5 ET provides higher clinical pregnancy outcomes with reduced risks of multiple pregnancies, it still has potential risks of developmental arrest of IVF embryos. The aim of this study was to evaluate the clinical outcomes of day 4 ETs and compare the efficacy of day 4 ET with day 5 ET. Methods: From 2006 to 2009, a total of 747 fresh IVF-ET cycles were retrospectively analyzed (day 4, n=440 or and day 5, n=307). The cycles with any genetic factors were excluded. The rates of matured oocytes, fertilization, good embryos, and clinical pregnancy of the two groups were compared. The chi-square test and t-test were used for statistical analysis. Results: There were no significant differences between the two groups with respect to the mean age of the females and rates of matured oocytes. The pregnancy outcomes of day 4 ET (40.7%) were similar to those of day 5 ET (44.6%). The implantation rate of day 5 ET (24.2%) was significantly higher than that of day 4 ET (18.4%) (p=0.003). Conclusion: Day 4 ET can be chosen to avoid ET cancellation in day 5 ET resulting from suboptimal circumstances in the IVF laboratory, but the decremented quality of embryos for transfer and the decreased pregnancy rate must be taken into consideration.

• 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.

• 한국수정란이식학회지
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• 제25권3호
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• pp.141-144
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• 2010

In vivo embryo produced from Hanwoo donor cows were collected and transferred to Hanwoo recipients. Cows, at random stages of the estrous cycle, received Progesterone Releasing Intravaginal Device (CIDR-plus, InterAg, New Zealand) together with injection of 1 mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment began 4 day later. For superovulation, a total of 28 mg FSH was intramuscularly injected twice a day in the way of decreasing doses 4 day (5, 5, 4, 4, 3, 3, 2 and 2 mg). Twenty one Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered 7 days after the second insemination by flushing the uterus with Embryo Collection Medium. The results obtained were as follows: The rates of transferable embryos were 50.3%, and 78 fresh embryos at morulae and blastocysts stage were transferred into Hanwoo recipients on day 7 of estrus cycle. The pregnancy rates were first embryo transfer 55.6%, 2nd 62.9% and 3rd 57.9%, respectively. In conclusion, These results suggest that CIDR-based superovulation protocol may be effectively used for production of superior Hanwoo embryos. Also, since it seems the condition of recipient cows greatly affect pregnancy rate, it is very important to evaluate recipient for effective cattle production.

• 한국수정란이식학회지
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• 제12권1호
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• pp.103-110
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• 1997

The present study was carried out to investigate effects of gonadotropin, age of donor, day of estrus cycle gonadotropin injection started and season on embryo production after superovulation in dairy cattles. Embryo collection records were obtained from 177 embryo donor collections from 98 Holstein cows aged from 3 to 9 years during 4 years(1993~1996) at National Livestock Re-search Institute. Superovulation was induced by injections of 3 gonadotropins(FSH-P, FOLLTROPHIN-v* or SUPER-OV*) beginning on days 9 to 14 of the estrus cycle. Em-bryos were collected from donors using a nonsurgical technique on days 6 to 8 after insemi-nation. The results were as follows ;Number of total and freezable embryos per donor cow was affected by gonadotropin(P<0.01). The more number of total and freezable embryos wereobtained by use of FOLLTROPHIN-V (13.2, 7.4) or FSH-P (11.0, 5.7) than SUPER-OV* (5.0, 2.4). Age of donor, the day that gonadotropin was started or season didn't affect total or freezable embryos(P >0.05).

• 한국수정란이식학회지
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• 제15권2호
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• pp.115-120
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• 2000

This study was undertaken to test the hypothesis the hypothesis that treatment with hCG (5,000 IU) at the time of embryo transfer would enhance pregnancy rates in recipients, and the concentration of plasma urea nitrogen(PUN) in recipients was related to the effect of hCG on the reproductive performance. Blood samples were taken according to experimental condition for the assessment of the endogenous plasma progesterone concentration and plasma urea nitrogen. Concentrations of progesterone in plasma were higher in cows treated with hCG on day 7(estrus=day 0) than in those untreated during 7∼43 days after insemination. The pregnancy rates were 65.5 and 54.6% for the hCG treated and untreated groups, respectively. In recipient group categorized with PUN concentration of <12 mg/이, the pregnancy rates were 68.8 and 46.7% for the hCG treated and untreated groups, respectively. The results suggest that hCG treatment at 7 days after insemination could be used to increase the pregnancy rate of embryo transfer, and transfer, and only the recipients with PUN concentration of <12 mg/dl were influenced by treatment with hCG.

• 한국수정란이식학회지
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• 제5권2호
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• pp.38-44
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• 1990

This study was carried out to produce superior dairy cattle by embryo transfer. Seven dairy cows were superovulated with divided injection of FSH 4Omg for 5 days started on day 9 to 14 of the estrus cycle and injection of PGF$_2$$\alpha$ 45mg on day 4 of FSH injection. Donor cows were flushed to collect embryos on day 7 or 8 of the estrus cycle. Fresh embryos collected were transferred to synchronized dairy recipients or frozen using glycerol 3 step method to he equilibrated. And 35 embryos which were frozen using glycerol 6 step method were imported from U.S.A. After glycerol dilution of frozen embryos was done by reverse density during freezing. frozen-thawed embryos were transferred to synchronized dairy or beef recipients. The results obtained were as follows; 1. Total of 24 embryos were collected from 7 donor cows flushed and transferable embryos were 18 (75.0%). 2. Among 24 embryos. morula, early blastocyst, blastocyst, expanded blastocyst and unfertilized ova were 3 (12.5%), 1 (4.2%), 10 (41.6%), 4 (16.7%) and 6 (25.0%), respectively. 3. Heat inducing rate after 1st and 2nd injections of PGF$_2$$\alpha$ in Holstein and beef cattle was 83.3% and 71.4% and 62.5% and 69.2%, respectively. 4. Among 56 recipients, 23 head were pregnant (41.1%). The pregnancy rate of fresh embryos was 50.0% (1/2 heads) and the pregnancy rate of frozen embryos which were frozen using glycerol 3 step and using glycerol 6 step imported from U.S.A. was 52.6%(l0/19 heads) and 34.3%(12/35 heads), respectively. 5. The pregnancy rate of blastocyst (60.0%) was higher than that of morula (39.0%), early blastocyst (25.0%) and expanded blastocyst (0%). 6. The pregnancy rate of grade I embryos (52.2%) was higher than that of grade 2 (34.6%) and grade 3 (28.6%). 7. The pregnancy rate according to synchrony of recipient with donor was higher in simultaneous recipient (55.0%) and +l2hrs' (53.8%) than -24hrs' (23.5%), -l2hrs' (20.0%) and +24hrs' (0%).

• 한국수정란이식학회지
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• 제4권1호
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• pp.41-45
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• 1989

Total thirty of flushing were attempted on day 4 to 15 of estrus cycle with S heifers and 9 cows by nonsurgical method. The flushed or recovered rate among flushings was 86.7% (26/30) or 88.5% (23/26), respectively. There was no difference in the recovered rate between heifers (85.7%,6/7) and cows (89.5%, 17119). The embryo was recovered on day 4 to 15 of estrus cycle from the donors in natural heat without any technical difficulties.The I2FG Foley catheter used for pubertal heifers had sometimes plug in it with uterine mucus during flushing of uterine horn. But the problem could be overcomed by pumping the catherter with fluthing solution or by changing the catheter. Three normal embryos were recovered from 3 pubertal (10-11 month old) heifers. The rate of normal and abnormal eggs was 60.9% (14123) and 39.1% (9/23), respectively. The abnormal eggs were on degenerating except one unfertilized egg and were mostly recovered from heifers or cows flushed consecutively during the estrus cycle. The developmental states of normal embryos were l6-cells on day 5, 32-cells on day 6, compacted-morula on day 7, early-to expanded-blastocyst on day 8-to 9, and hatching-to hatched-blastocyst on day 10 to 11 of estrus cycle. The stage of embryos on day 8 to 10 showed varities among donors. On day 8 to 9 of estrus cycle hatching-blas tocyst was recovered from some donors.

• 한국수정란이식학회지
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• 제5권2호
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• pp.1-10
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• 1990

The individual difference of superovulatory responses and inferior embryo quality in superovulated cattle may cause disturbances in the endocrine profile, follicular steroidogenesis, nuclear maturation of nocyte, fertilization and cleavage of embryos. However, the reasons why those disturbances are occurred were not understood. The methods of the improvement of superovulatory response and embryo production were the use of anti-PMSG if PMSG used, pure FSH or controlled FSH-LH inducer, priming dose of gonadotropin in the first few day of the estrous cycle and GnRH or analogue. However, all of the above methods were not reduced the individual differences but improved embryo production We must continue the fundamental studies to understand the mechanism.

• 한국수정란이식학회지
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• 제17권1호
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• pp.23-32
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• 2002

본 연구는 기존의 수정란 이식기술을 다각적으로 분석하고 개선하여 한우 체내수정란의 생산 및 이식기술 체계를 확립하기 위하여 수행하였으며, 결과를 요약하면 다음과 같다. 1. 배란에 미치는 FSH analogue 간의 유의성은 인정되지 않았으나 수정률, 이식 가능 수정란 및 동결수정란 생산에는 처리간에 유의성(P<0.05)이 인정되었다. 특히, Super-OV는 Foll-tropin-V 및 Embryo-S에 비해서 이식 가능 수정란의 생산율이 유의적(P<0.01)으로 낮았다. 2. 호르몬의 투여시기에 따라 공란우의 배란정도 (P<0.1), 수정률, 이식 가능 및 동결 가능 수정란의 생산(P<0.05)은 각각 유의적인 차이를 나타냈다. 특히, 발정주기 11일차의 호르몬투여는 적합하지 않은 결과를 보였다. 3. 다배란처리후 황체의 등급에 따른 수정란의 수는 좌측 또는 우측에서 6개 이상의 황체를 보인 A등급(8.06∼9.49개)의 경우가 B, C 및 D등급(2.93 ∼6.28개)보다 유의적(P<0.01)으로 높았다