• Reproductive and Developmental Biology
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• v.28 no.4
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• pp.221-226
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• 2004

The present study was performed to investigate the effects of in vitro maturation (IVM) and in vitro fertilization (IVF) duration on the development of Korean Native Cattle embryos. The time of blastocyst formation and the quality of blastocysts based on cell numbers were examined. The cleavage rate increased with the length of IVF duration in the groups of 18-hr IVM, but was constant in the groups of 24-hr IVM. The development rate to the 8-cell stage was significantly higher in the IVM 18: IVF 20 group than in the IVM 24: IVF 24 group. The development rate to the blastocyst stage was highest in the IVM 18: IVF 20 group, significantly different from that of the IVM 18: IVF 16, IVM 24: IVF 20 and IVM 24: IVF 24 group. The time of blastocysts formation tended to be shorter when IVM and IVF duration were decreased. The number of inner cell mass, trophoblast and the total cells were significantly higher in the IVM 18: IVF 16 group than in the IVM 24: IVF 24 group (P<0.05). These results demonstrated that the IVM and IVF duration should be adequate for the efficient production of bovine embryos, and it might particularly be essential to determine the proper combination of IVM and IVF duration.

• Korean Journal of Animal Reproduction
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• v.14 no.1
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• pp.17-22
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• 1990

The present study was carried to investigate the development potential of bovine follicular oocytes matured and fertilized in vitro. Bovine oocytes matured in vitro were fertilized, cultured, and transfered to the rabbit oviduct for in vitro culture to evaluate the development potential. The rates of in vitro maturation, fertilization and polyspermy were 96%(355/369), 90%(320/355) and 15%(17/320), respectively. The percentage of oocytes cleaved after culture for 48 hours, to 2 cell, 3~4 cell, 6~8 cell stage were 17%(60/356), 21%(75/356) and 19%(67/356), respectively and overall cleavage rate was 57%(202/356). The rate of recovered embryos after 5 days in rabbit oviduct was 56%(80/142), and 21%(17/80) of recovered embryos developed over morula stage.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.8
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• pp.1117-1123
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• 2009

This study investigated the effects of IGF-I and EGF on the development of blastocysts or hatched blastocysts during the in vitro culture of embryos from immature porcine oocytes. After the in vitro maturation and fertilization of cumulus-oocyte complexes (COCs) and their culture in vitro in PZM3 medium, we examined the embryo development rate for 168 h. When different concentrations of IGF-I (0, 1, 10, 20 ng/ml) were supplemented to fertilized porcine embryos in vitro, there were no significant differences in cleavage rate, blastocyst development rate or blastocyst hatching rate among the treated groups. On the other hand, when different concentrations of EGF (0, 1, 10, 20 ng/ml) were supplemented to the in vitro culture medium, blastocyst development rate was highest in the group in which EGF was not supplemented and, specifically, it was higher than in the 20 ng/ml treatment group (p<0.05). When 10 ng/ml IGF-I and 1 ng/ml EGF were supplemented separately or simultaneously, there were no significant differences among the treated groups in blastocyst hatching rate and the number of cells in each condition. This study demonstrated that the addition of IGF-I and EGF into PZM3 medium did not enhance development of the blastocyst stage and total cell number in blastocysts.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.170-170
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• 2004

First of all, in vitro production (IVP) of porcine embryos is an important as initial step to improve bio-technical applications such as transgenesis and cloning for xenotransplantation. In recent years, considerable progress has been achieved in the IVP embryos using advanced methods for in vitro maturation (IVM) and fertilization (IVF). (omitted)

• Korean Journal of Animal Reproduction
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• v.19 no.1
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• pp.9-14
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• 1995

This study was conducted to investigate the effect of energy source on development of in vitro development of in vitro matured and fertilized porcine 2-cell embryos. The relative preferences of glucose, lactate and pyruvate for in vitro development of porcine 2-cell embryos were determined. The results obtained are as follows. 1. 33.3, 20.8 and 29.2% of porcine embryos reached morula stage in addition to lactate, glucose, and both glucose and lactate in the culture medium as energy source, respectively. 2. 38.5, 15.4 and 26.9% of porcine embryos reached morula stage in addition to pyruvate, glucose, and both glucose and pyruvate in culture medium as energy source, respectively. 3. 42.9, 21.4 and 28.6% of porcine embryos reached morula stage in addition to pyruvate and lactate, glucse alone, and glucose, lactate and pyruvate in culture medium as energy source, respectively.

• Reproductive and Developmental Biology
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• v.28 no.2
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• pp.101-105
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• 2004

This study was carried out to evaluate the effects of glucose and sodium phosphate on in vitro development of porcine oocytes matured and fertilized in vitro. When the culture medium was supplemented with various concentrations of glucose, the higher proportions (23 and 26%) of oocytes developed to morular or blastocyst stages were at the concentrations of 2.78 and 5.56 mM than 0 (9%; P<0.05) and 11.12 mM (18%). In experiment to evaluate effect of sodium phosphate during in vitro development of porcine oocytes, a significantly (P<0.05) higher proportions of embryos developed to morular or blastocyst stages was obtained with sodium phosphateof 0.28 (25%) and 0.53 (27％) mM than 0 (15%), 1.05 (19%) and 2.10 (10%) mM. On the other hand, when oocytes were cultured in medium with (0.53 mM) sodium phosphate, the proportions of developed embryos were significantly (P<0.05) higher in medium without (29%) that than with (14%) 5.56 mM glucose. However, a higher proportion of embryos developed to morular or blastocyst stages were obtained in medium with (23%) that than without (8%) glucose (P<0.05). The minimum essential medium (MEM) added to the culture medium were higher regardless of presence of sodium phosphate and glucose on the development of embryos. Although sodium phosphate and glucose could support morular and blastocyst development to a limited extend (10∼24%), significantly higher proportion (36%) at morular or blastocyst stages was obtained by MEM adding in the medium with sodium phosphate and glucose. These results suggest that the early development of in vitro fertilized porcine oocytes can be maintained efficiently by glucose and sodium phosphate when they were cultured in medium with MEM.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.2
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• pp.117-123
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• 1992

Development in vitro of 2-cell mouse embryos was examined after appropriate exposure to oviductal milieu to demonstrate biological activity present in the oviducts. ICR and ($C57Bl/6{\times}Balb/c$) $F_1$ hybrid mice were superovulated and mated for the recovery of early embryos. Embryos were recoverd at every 2h intervals from 32h post-hCG(hph) to 56 hph. The proportions of developmental stages were determined in the recovered embryos. Development in vitro of 2-cell embryos was more rapid in $F_1$ hybrid than in ICR, showing high proportions of 4-cell embryo and blastocyst at 120 hph. 100% of blastocyst development was obtained at 38hph in $F_1$ hybrid and at 50 hph in ICR when 2-cell embryos were cultured upto 120hph in vitro. Moreover, in vitro culture of oviducts containing 2-cell embryos in ICR mice for 12h from 34hph to 46hph increased developmental capacity of ICR mouse embryo in vitro. The results indicate that oviductal environment contains substances having mitogenic activity and overcoming early cell block in vitro. The mitogenic activity is effective in vitro as well as in vivo.

• Journal of Veterinary Clinics
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• v.36 no.5
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• pp.253-258
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• 2019

The objective of this study was to analyse the effects of MS-275 (Class I and II histone deacetylase inhibitor) supplementation on the development of porcine in-vitro somatic nuclear transfer embryo production. During in-vitro development, early embryos were exposed to different concentrations of MS-275 (0, $5{\mu}M$, $10{\mu}M$, and $20{\mu}M$). In in-vitro culture supplemented group, the blastocyst development rate was significantly enhanced by $10{\mu}M$ concentration than other groups (24.0% vs. 19.3%, 21.8%, 11.5%; P < 0.05). Additionally, the 6 h supplementation group, significantly improved the blastocysts production than 24 h, 48 h and control groups (26.1% vs. 17.0%, 15.2%, 2.8%; P < 0.05). Following supplementation with optimal concentrations and time ($10{\mu}M$-6 h group), the blastocyst production was significantly higher than control (25.7% vs 15.8%; P < 0.05). The optimal concentrations of MS-275 significantly enhanced the percentages of ICM:TE than control (43.6% vs. 38.4%; P < 0.05) accompanied with significantly higher expression levels of reprogramming related genes (POU5F1, Naong, and SOX2). In conclusion, the optimal concentrations of $10{\mu}M$ MS-275 and 6 h supplementation during in-vitro culture can significantly improve the quality of porcine in-vitro somatic nuclear transfer embryos through histone acetylation and epigenetic modification. Increasing the efficiency of clonal animal production will greatly promote the development of animal disease models and xenotransplantation.

• Journal of Embryo Transfer
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• v.14 no.3
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• pp.219-224
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• 1999

The effect of several potential antioxidants were examined as a means of increasing the in vitro development of in vitro matured and in vitro fertilized oocytes into morulae and blastocysts. Korean native cattle embryos after in vitro fertilization were cultrued for 7 days at 38.5$^{\circ}C$ in CR1aa containing varing concentration of the antioxidants in a gas phases consisting of 5% CO2, 95% humidified air. The results obtained were summarized as follows; The proportion of embryos developed to morulae and blastocysts in CR1aa containing 2.5uM $\alpha$-tocopherol(11.0% and 6.0%) was significantly higher than those of 0, 5.0, and 7.5uM $\alpha$-tocopherol (P<0.05). concentration of 50uM L-ascorbic acid (7.5% blastocysts) did affect the proportion of embryos developing into blastocystes(P>0.05). Addition of 200uM cysteamine was significantly higher than those of 0, 100 and 300uM (P<0.05). When the fertilized oocytes were cultured at 0. 200, 400 and 600uM of selenium for 168 hrs, the morulae rates were 12.2, 5.2, 16.0 and 16.1% respectively, and addition of 200uM selenium was significantly higher than those of 0, 400, 600uM (P<0.05). These results suggested that the addition of $\alpha$-tocopherol, L-ascorbic acid, cysteamine and selenicum can enhanced development to the morulae and blastocysts of in vitro derived fertilized oocytes.

• Journal of Embryo Transfer
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• v.26 no.2
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• pp.117-122
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• 2011

In this study, we examined the effectiveness of in vitro fertilization of porcine immature oocytes on the embryo development of blastocysts or hatched blastocysts and the number of cells according to the in vitro fertilization conditions. In the in vitro fertilization of in vitro matured porcine oocytes, there were no significant differences between treatment groups regarding fertilization rate, blastocyst rate, and embryo development of hatched blastocysts according to the storage periods of liquid sperm of 24, 48, and 72 hours. The embryo development rate of hatched blastocysts after the fertilization according to different spermatozoa concentrations ($0.4{\times}10^5$, $1.2{\times}10^5$, and $3.6{\times}10^5$ cells/ml) showed the highest rate in the group with a spermatozoa concentration of $1.2{\times}10^5$ cells/ml; in particular, this rate was significantly higher than that in the $0.4{\times}10^5$ cells/ml group (p<0.05). The total number of blastocysts cells as well as trophectoderms (TE) that developed in each treatment group were also significantly higher in the $1.2{\times}10^5$ cells/ml group than in any other groups (p<0.05). In contrast, the embryo development rate of blastocysts according to different co-incubation periods of sperm and oocyte (1, 3, and 6 hr) was high in the 6-hour group; in particular, the rate was significantly higher than that of the I-hour group (p<0.05). Furthermore, the total number of oocytes cells and TEs that developed was significantly higher in the 6-hour group than any other group (p<0.05). In this study, the most effective treatment conditions for porcine embryo development and high cell number were found to be as follows: a sperm storage period of less than 72 hours, a spermatozoa concentration of $1.2{\times}10^5$ cells/ml, and a 6-hour co-incubation period for sperm and ooocyte.