• 한국수정란이식학회지
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• 제22권4호
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• pp.209-212
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• 2007

This study was performed to establish the early pregnancy diagnosis with evaluation of size of embryonic vesicles and change of crown-rump length (CRL) by transrectal ultrasonography in the Korean black goats. Early pregnancy diagnosis was performed in the 114 goats from 19 to 35 days after coitus by transrectal 5.0 MHz transducer. Embryonic vesicles and CRL in the 142 goats from 19 to 42 days was measured to establish the correlation between gestational age and these parameters. The early pregnancy diagnosis achieved 80% accuracy after days 25 and 100% accuracy after days 28 after breeding. Diameters of embryonic vesicle and CRL were significantly increased with the gestational age. In these facts, early pregnancy diagnosis is possible on Days 25 after breeding and measurement of embryonic vesicles and CRL were useful to predict the gestational age in Korean black goat.

• Asian-Australasian Journal of Animal Sciences
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• 제14권9호
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• pp.1342-1351
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• 2001

Over the years, the embryo transfer industry has grown from the simple collection & transfer of embryos into an advanced field of embryo biotechnology. Currently a large demand exists for bovine oocytes and early embryos in both research and commercial settings. Bovine embryos can now be produced in-vitro. Primary oocytes collected from antral follicles of abattoir - obtained ovaries can be induced to undergo the maturation process. In-vitor maturation system, however must ensure that the resulting oocyte is capable of undergoing normal fertilization and yields a zygote competent of developing to term after embryo transfer. Sperm preparation for IVF has improved with the use of heparine. The use of co-culture system has proved beneficial in circumventing the developmental block in IVM/IVF bovine embryos.

• 한국동물생명공학회지
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• 제35권3호
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• pp.215-222
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• 2020

Mesenchymal stem cells (MSCs) have been widely used as donor cells for somatic cell nuclear transfer (SCNT) to increase the efficiency of embryo cloning. Since replicative senescence reduces the efficiency of embryo cloning in MSCs during in vitro expansion, transfection of telomerase reverse transcriptase (TERT) into MSCs has been used to suppress the replicative senescence. Here, TERT-transfected MSCs in comparison with early passage MSCs (eMSCs) and sham-transfected MSCs (sMSCs) were used to evaluate the effects of embryo cloning with SCNT in a porcine model. Cloned embryos from tMSC, eMSC, and sMSC groups were indistinguishable in their fusion rate, cleavage rate, total cell number, and gene expression levels of OCT4, SOX2 and NANOG during the blastocyst stage. The blastocyst formation rates of tMSC and sMSC groups were comparable but significantly lower than that of the eMSC group (p < 0.05). In contrast, tMSC and eMSC groups demonstrated significantly reduced apoptotic incidence (p < 0.05), and decreased BAX but increased BCL2 expression in the blastocyst stage compared to the sMSC group (p < 0.05). Therefore, MSCs transfected with telomerase reverse transcriptase do not affect the overall development of the cloned embryos in porcine SCNT, but enables to maintain embryo quality, similar to apoptotic events in SCNT embryos typically achieved by an early passage MSC. This finding offers a bioengineering strategy in improving the porcine cloned embryo quality.

• 한국수정란이식학회지
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• 제10권3호
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• pp.251-256
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• 1995

This study was experimented that developmental effects of bovine in vitro fertilized embryos by coculture system and supplementation of energy materials into simple media. With the ovaries from slaughter house in vitro maturation by 24h, in vitro fertilization was performed with sperms collected by Percoll gradient method. Fertilized embryos were cocultured in 15% FCS+CZB medium with BOEC(bovine oviductal epithelial cell), GCM (granulosa cell monolayer) and MEFC(mouse embryonic fihrohlast cell). And also in this study, there was trying to improve the early developmental rate of embryos by addition of concentration-controlled Na-pyruvate, D-glucose which were used as energy sources into CZB medium. In vitro developmental rate was confirmed by the cleavage rate of 48h post-IVF and the embryo development rate at 240h culture. In the coculture system BOEC had 20.0% of blastocysts rate, which was higher than that of other coculture systems. To determine the optimum concentration for early embryo developmental rate rapidly, through the gradient of concentrations of Na-pyruvate and D-glucose, we focused on the cleavage rate at 48h and blastocysts rate at 240h. In case of Na-pyruvate, cleavage rate and developmental rate over 3-cell were lower at the concentration of 1.OOrnM than the other treatment concentrations, otherwise the blastocysts rate was higher as 23.2% than the others. That result showed that as like reported group which had higher develop-mental rate over 3-cell was also higher to the blastocysts rate. In case of D-glucose, there was no effects through the concentration changes. It was the result of this study for which the use of BOEC coculture system and 1.OOmM Na-pyruvate as an energy source had an effect upon embryo development.

• 한국수정란이식학회지
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• 제31권3호
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• pp.215-219
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• 2016

Value of excellent breeding animals is important in livestock industry, but their economic life time is limited. And, many countries have been trying procuration of genetic resource in good animals. Therefore, this study was conducted to determine embryo production and to test efficiency of embryo transfer via non-surgical artificial insemination (AI) in different breed of superior sows. A total of 17 sows were used in this experiment (Duroc, n=10; Landrace, n=4; Yorkshire, n=3). The sows were artificially inseminated by semen of same breed boars. After 4 or 5 days following the AI, the embryos were obtained from the sows and then transferred to Landrace and Yorkshire recipients (n=3, respectively) by non-surgical method. The corpora lutea tended to be increased in Yorkshire and Landrace than Duroc(28 and 26 vs. 17, respectively). The recovery of embryo was 78.8% in Landrace, 65.4% in Duroc and 51.4% in Yorkshire. Duroc showed lower morulaes and early blastocyst embryos than 2, 4, 8 and 16 cell. The morula in Yorkshire was higher (P<0.05) than that of Duroc (4.7 vs. 3.4). Similarly, the morulaes and early blastocyst embryos presented greater (P<0.05) in Landrace compared with other breed sows. The recipient sows were pregnant in a Landrace only. This reason may be due to little embryos inserted in the recipients. In addition, pregnancy results were limited because of the little sows. In conclusion, ovulated ovum in sows can be affected by different breed. Also, further study needed pregnant test by using the many embryo in each breed.

• 한국수정란이식학회지
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• 제6권2호
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• pp.1-17
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• 1991

It is widely recognized that the embryonic or fetal loss after breeding is common in the cattle and that it is an important factor affecting reproductive efficiency. The causes of this loss have been subject of extensive researches and the results indicate that the embryonic mortality may he primary factor responsible for low pregnancy rates in non-embryo transfer bovine populations as well as embryo transfer programs. However, it's causes are still not clearly understood. The embryonic mortality or pregnancy rate has been influenced by various embryonic and maternal effects related to genetic and environmental factors. The timing and extent of embryonic mortality vanes greatly according to authors and estimating methods, because it is difficult to make direct measurements. The major important factors that may influence the embryonic losses or pregnancy rates after embryo transfer can be summeirized. 1.When an embryo is transferred to unmated recipients, the contralateral transfer to corpus luteum results in a lower survival rate than ipsilateral deposition. When the embryos are transferred for the production of twin calves, their survivals and twin pregnancies have quite inconsistent according to the transfer methods either to the unmated-synchronized or already mated recipients and more works are needed to accurrately clarify the previous results. 2.Although embryos can be cultured in vitro some hours without the great declines in pregnancy rates, the rates differ markedly among culture times and media but may be improved by co-transfer systems. 3.Embryo developmental stages and quality grades clearly affect the survival rate following freezing and the pregnancy rate after transfer and the selection of embryos without chromosome abnormalities and of high fertile semen may also be considered to increase the pregnancy rates. 4.Many researches have attempted to relate the plasma progesterone levels to pregnancy rates and others have done either direct progesterone supplementation or luteal stimulation by hCG treatment in order to increase the pregnancy rates. However, these effects on pregnancy rates are inconsistent and also contradictory. 5.The asynchrony between donors or embryos and recipients may he a major cause of embryo death and low pregnancy rate and the sensitivity to uterine asynchyony differs in according to the quality and stages of embryos. 6.The extremes of poor or over nutrition during early pregnancy in the recipients are detrimental to the survival of embryos and the good body condition is required to prevent a reduejion of pregnancy rates. The uterine pathogens in embryonic mortality or fertility have been questioned but the infection of C.pyogenes and Campylobacter fetus is still important pathogens. 7.The heat stress during early pregnancy may reduce conceptus weight and possibly increase the embryonic mortality.

• 한국수정란이식학회지
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• 제11권3호
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• pp.201-210
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• 1996

This experiment was carried out to evaluate the effect of early mouse embryonic development in vitro by co-culture with bovine and porcine oviductal epithelial cells (BOEC and POEC). The 2-cell embryos were collected from the oviducts of the superovulated and mated cultured in D-PBS /15% FCS at 48 hours after hCG injection. The in vitro developmental rate of blastocyst formation in the embryos were examined under the fllowing treatments; 1) TCM 199 added 15% HCS, 2) Ham's F-10 added 15% HCS, 3) MediCult IVF medium, 4) TCM 199 added 15% HCS + BOEC, 5) TCM 199 added 15% HCS + POEC, 6) Ham's F40 added 15% HCS + BOEC, 7) Ham's F-10 added 15% HCS + POEC,8) MediCult IVF medium + BOEC, 9) MediCult IVF medium + POEC. For a comparative study of in vitro development for 96 hours after hCG injection, were cultured with oviductal epithelial cell and media only. The obtained results were 2-cell embryos developed to the blastocyst stage in TCM 199, Ham's F-10 and MediCult IVF medium at the rates of 84.4,83.2 and 81.6%. respectively. The higher developmental rates(91~97%) of blastocyst formation was appeared when the embryos were co-cultured with a monolayer of bovine or porcine oviductal epithelial cells in TCM 199 or Ham's F-10 and MediCult IVF media. No significant difference in developmental rates was shown between bovine and porcine oviductal epithelial cells but significant difference in co-culture system in comparison between media only system and co-cultures. In conclusions, oviductal epithelial cells, BOEC and POEC, when co-culture with mouse early embryos improved the rates of development, blastocyst and hatching. Therefore, it is suggested that co-culture system using oviductal epithelial cells improve early embryonic developtnent in mouse.

• Asian-Australasian Journal of Animal Sciences
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• 제25권12호
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• pp.1681-1690
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• 2012

The aim of this study was to investigate the impact of a reported p53 inhibitor, pifithrin-${\alpha}$ (PFT-${\alpha}$), on preimplantation porcine in vitro fertilized (IVF) embryo development in culture. Treatment of PFT-${\alpha}$ was administered at both early (0 to 48 hpi), and later stages (48 to 168 hpi) of preimplantation development, and its impact upon the expression of five genes related to apoptosis (p53, bak, bcl-xL, p66Shc and caspase3), was assessed in resulting d 7 blastocysts, using real-time quantitative PCR. Total cell numbers, along with the number of apoptotic nuclei, as detected by the in situ cell death detection assay, were also calculated on d 7 in treated and non-treated control embryos. The results indicate that PFT-${\alpha}$, when administered at both early and later stages of porcine IVF embryo development, increases the incidence of apoptosis in resulting blastocysts. When administered at early cleavage stages, PFT-${\alpha}$ treatment was shown to reduce the developmental competence of porcine IVF embryos, as well as reducing the quality of resulting blastocysts in terms of overall cell numbers. In contrast, at later stages, PFT-${\alpha}$ administration resulted in marginally increased blastocyst development rates amongst treated embryos, but did not affect cell numbers. However, PFT-${\alpha}$ treatment induced apoptosis and apoptotic related gene expression, in all treated embryos, irrespective of the timing of treatment. Our results indicate that PFT-${\alpha}$ may severely compromise the developmental potential of porcine IVF embryos, and is a potent apoptotic agent when placed into porcine embryo culture media. Thus, caution should be exercised when using PFT-${\alpha}$ as a specific inhibitor of p53 mediated apoptosis, in the context of porcine IVF embryo culture systems.

• 한국수정란이식학회지
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• 제14권1호
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• pp.1-8
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• 1999

The efficiency of transgenic livestock animal production may be improved by early selection of transgenci preimplantation embryos. To examine the possibility of GFP gene as a non-invasive marker for the early screening of transgenic embryo, the GFP gene was microinjected into rabbit zygotes and the later stages of preimplantation embryos were examined for the expression of GFP. The presence of injected DNA was detected by PCR analysis and the expression of GFP was detected by observing green fluorescence in embryos under a fluorescent microscope. Out of 108 GFP gene-injected rabbit zygotes, seventy three(67.6%) were fluorescence-positive. When 11 fluroresecence-positive blastocysts were analyzed for the presence of GFP gene by PCR, 6(54.5%) were positive, and all of the 8 flrouescence-negative blastocysts were also negative by PCR. The results indicate that the screening of transgene in rabbit embryos by PCR analysis and GFP detection could be a promising method for the preselection of transgenic embryos.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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• pp.87-87
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• 2003

Taurine (2-aminoethanesulfonic acid, $^{+}NH_3CH_2CH_2{SO_3}^{-}$) is endogenous $\beta$-amino acid which is essential in fetal nutrition and development and is present in abundant quantities in several tissues of fetus. In utero, taurine deficiency causes abnormal development and abnormal function of brain, retina, kidney and myocardium. Thus, transfer of taurine into fetus is important during embryonic development. Taurine transporter (TauT) has 12 hydrophobic membrane -spanning domains, which is typical of the $Na^{+}$- and $Cl^{-}$-dependent transporter gene family. Among the various biosynthetic enzymes of taurine, cysteine sulfinic acid decarboxylase (CSD) is the rate-limiting enzyme for biosynthesis of taurine. However, the enzyme activities of taurine biosynthesis are limited in early stage of embryonic development. To analyze the expression period of TauT and CSD during embryonic development, we have investigated the gene expression of TauT and CSD using reverse transcriptase polymerase chain reaction (RT-PCR) in mouse and chicken embryos. RT-PCR anaylsis revealed that both TauT and CSD mRNAs were already expressed at Day-4.5 in mouse embryo. In chicken whole embryo, TauT and CSD mRNAs began to appear on developing times of 48 hrs and 12 hrs, respectively. TauT mRNA was detected in the organs of heart, brain and eye of the day-3 chicken embryo. Our data show that TauT and CSD mRNAs were expressed in early stage of embryonic development.