• Proceedings of the Korean Society of Developmental Biology Conference
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• 2004.07a
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• pp.24-24
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• 2004

• Reproductive and Developmental Biology
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• v.28 no.3
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• pp.141-145
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• 2004

Mucin coat is deposited on the embryos during passage through the oviduct in rabbit. When in vitro cultured blastocysts were transferred to the recipients, the lack of mucin coat might account in part for failure of pregnancy after transfer. The present study were carried out to investigate whether deposition of mucin coat were induced when in vitro cultured blastocysts were transferred to recipients. At 19 ～20 hours post-coitus one-cell embryos were collected by flushing oviducts. These embryos cultured for 72 hours were reached to blastocyst stage. And these blastocysts were transferred to the oviduct of asynchronized (one day later than the donors) and synchronized recipient. To confirm deposition of the mucin coat, blastocysts transferred to the oviduct were recovered at 24 and 48 hours after the transfer. Fifty eight percent of blastocysts recovered from uterus of asynchronous recipient at 24 hours after transfer and 92.9% of blastocysts recovered from uterus of synchronous recipient were 0～10 ㎛ of mucin coat thickness. And 11.8% of blastocysts of asynchronized recipients and 7.1% of blastocysts from asynchronized recipients were in 11～20 ㎛ of mucin coat thickness. When blastocysts were recovered from uterus at 48 hours after transfer, 87.0% of blastocysts from asynchronized recipients and 5.9% of blastocyst from synchronized recipients were in 0～10 ㎛ of mucin coat thickness. And 76.5% of blastocysts of synchronized recipients and 4.4% of blastocysts from asynchronized recipients were in 11～20 ㎛ of mucin coat thickness. From these results it is speculated that the low implantation rate of in vitro cultured rabbit blastocysts transferred to oviduct of recipient was caused by high degeneration of the embryo after transfer and inappropriate deposition of mucin coat.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.585-592
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• 2017

Objective: The present study investigates pre- and post-implantation developmental competence of nuclear-transferred porcine embryos derived from male and female fetal fibroblasts. Methods: Male and female fetal fibroblasts were transferred to in vitro-matured enucleated oocytes and in vitro and in vivo developmental competence of reconstructed embryos was investigated. And, a total of 6,789 female fibroblast nuclear-transferred embryos were surgically transferred into 41 surrogate gilts and 4,746 male fibroblast nuclear-transferred embryos were surgically transferred into 25 surrogate gilts. Results: The competence to develop into blastocysts was not significantly different between the sexes. The mean cell number of female and male cloned blastocysts obtained by in vivo culture ($143.8{\pm}10.5$ to $159.2{\pm}14.8$) was higher than that of in vitro culture of somatic cell nuclear transfer (SCNT) groups ($31.4{\pm}8.3$ to $33.4{\pm}11.1$). After embryo transfer, 5 pregnant gilts from each treatment delivered 15 female and 22 male piglets. The average birth weight of the cloned piglets, gestation length, and the postnatal survival rates were not significantly different (p<0.05) between sexes. Conclusion: The present study found that the sex difference of the nuclear donor does not affect the developmental rate of porcine SCNT embryos. Furthermore, postnatal survivability of the cloned piglets was not affected by the sex of the donor cell.

• Reproductive and Developmental Biology
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• v.36 no.4
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• pp.249-254
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• 2012

This study aimed at investigating whether a porcine follicular fluid (pFF) supplementation positively affects the characteristics of donor cells and the developmental competence of porcine cloned embryos. Ear fibroblast cells (donor cell) from an Massachusetts General Hospital miniature pig were cultured in different culture methods: (1) Dulbecco's modified Eagle's medium (DMEM)+10% FBS (Control); (2) DMEM+0.5% FBS (SS); and (3) DMEM+10% FBS+10% pFF (pFF) for 72 h. In each conditioned medium, the concentrations of 4 amino acids (Thr, Glu, Pro, and Val) in the pFF group were significantly different from those in the control group (p<0.05 or p<0.01). The proliferation of the cells cultured in the SS group was significantly lower than that of the other treatment groups (p<0.01). The population of apoptotic and necrotic cells in the SS group was significantly higher than that of either the control or the pFF group (p<0.01). The number of embryos that cleaved (p<0.05) and developed into blastocysts (p<0.01) in the SS group was significantly lower than that of either the control or the pFF group. Compared to other groups, the blastocysts produced from the donor cells in the pFF group had higher total cells and lower apoptotic cells (p<0.05). It can be concluded that pFF supplementation in the donor cell culture medium positively affects cell death, cell cycle and quality of the cloned blastocyst.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.9
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• pp.1255-1261
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• 2013

Somatic cell nuclear transfer (SCNT) is a useful tool for animal cloning, but the efficiency of producing viable offspring by SCNT is very low. To improve this efficiency in the production of cloned pigs, it is critical to understand the interactions between uterine function and cloned embryos during implantation. Lysophosphatidic acid (LPA) is a lipid mediator that plays an important role in the establishment of pregnancy in pigs; however, LPA production in the uterine endometrium of pigs carrying SCNT-cloned conceptuses has not been determined. Therefore, we investigated expression of ENPP2, an LPA-generating enzyme, in the uterine endometrium of gilts with conceptuses derived from SCNT during the implantation period. Uterine endometrial tissue and uterine flushing were obtained from gilts carrying SCNT-derived conceptuses and from gilts carrying conceptuses resulting from natural mating on d 12 of pregnancy. Our results demonstrated no difference in the level of ENPP2 mRNA expression in the uterine endometrium between gilts carrying SCNT-derived conceptuses and gilts carrying naturally-conceived conceptuses, but secretion of ENPP2 protein into the uterine lumen did decrease significantly in pigs with SCNT-derived conceptuses. These results indicate that expression and secretion of ENPP2, which are critical for appropriate LPA production and successful pregnancy, are dysregulated in the uterine endometrium of pigs carrying SCNT-derived conceptuses.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.73-73
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• 2002

Nuclear transfer (NT) techniques have advanced in the last years, and cloned animals have been produced by using somatic cells in several species including pig. However, it is difficult that the nuclear transfer porcine embryos development to blastocyst stage overcoming the cell block in vitro. Abnormal segregation of chromosomes in nuclear transferred embryos on genome activation stage bring about embryo degeneration, abnormal blastocyst, delayed and low embryo development. Thus, we are evaluated that the correlations of the frequency of embryo developmental rates and chromosome aberration in NT and In viかo fertilization (IVF) derived embryo. We are used for ear-skin-fibroblast cell in NT. If only karyotyping of embryonic cells are chromosomally abnormal, they may difficultly remain undetected. Then, we evaluate the chromosome aberrations, fluorescent in situ hybridization (FISH) with porcine chromosome 1 submetacentric specific DNA probe were excuted. In normal diploid cell nucleus, two hybridization signal was detected. In contrast, abnormal cell figured one or three over signals. The developmental rates of NT and IVF embryos were 55% vs 63%, 32% vs 33% and 13% vs 17% in 2 cell, 8 cell and blastocyst, respectively. When looking at the types of chromosome aberration, the detection of aneuploidy at Day 3 on the embryo culture. The percentage of chromosome aneuploidy of NT and IVF at 4-cell stage 40.0%, 31.3%, respectively. This result indicate that chromosomal abnormalities are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT associated with lower implantation rate, increase abortion rate and production of abnormal fetuses.

• Korean Journal of Agricultural Science
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• v.44 no.3
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• pp.309-317
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• 2017

Melatonin (N-acetyl-5-methoxytryptamine) is an indole synthesized from tryptophan by the pineal gland in animal. The major function of melatonin is to modulate circadian and circannual rhythms in photoperiodic mammals. Importantly, however, melatonin is also a free radical scavenger, anti-oxidant, and anti-apoptotic agent. Recently, the beneficial effects of melatonin on oocyte maturation and embryonic development in vitro have been reported in many species such as pig, cattle, sheep, mouse, and human. In this review, we will discuss recent studies about the role of melatonin in the production of porcine and bovine oocytes and embryos in vitro in order to provide useful information of melatonin in oocyte maturation and embryo culture in vitro.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.10a
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• pp.25-31
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• 2001

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.136-136
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• 2003

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.124-124
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• 2003