• Clinical and Experimental Reproductive Medicine
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• v.34 no.4
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• pp.253-273
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• 2007

Objective: This study was to find ways to let a manager or superintendent rationally and consistently inspect as well as let a embryologist precisely record [The Book of Supernumerary Embryo Preservation] and [The Book of Supernumerary Embryo Donation]. Methods: Based on the data collected between 1994 and 2004 in Clinic 44 (Maria Fertility Hospital), [The Present State about Production and Use of Embryos], [The Preservation of Supernumerary Embryos and Their Thaw State], [The Present State about Thaw and Use of Frozen Embryos], [The Present State about Donation and Charge of Frozen Embryos], [The Book about Frozen Embryo Discard], and [The Summarization Book about Management and Use of Frozen Embryos] were designed and recorded. Results: The production, use, preservation, discard and donation quantity of human embryos, the use and discard quantity of thawed embryos, and the cumulative embryo preservation quantity could be totalized in [The Present State about Production and Use of Embryos in Clinic 44]. Also, [The Preservation of Supernumerary Embryos and Their Thaw State in Clinic 44] supported "the supernumerary embryo preservation quantity" etc. In addition, [The Present State about Thaw and Use of Frozen Embryos in Clinic 44] or [The Book about Frozen Embryo Discard in Clinic 44] supported "the use and discard quantity of thawed embryos" etc. Moreover, "The embryo donation quantity" could be totalized in [The Present State about Donation and Charge of Frozen Embryos in Clinic 44]. Finally, [The Summarization Book about Management and Use of Frozen Embryos in Clinic 44] could be used for rational and consistent management or inspection. Conclusion: The present results suggest that the documents not only be standard data to record [The Book about Supernumerary Embryo Preservation in Clinic] and [The Book about Supernumerary Embryo Donation in Clinic] but can also be preserved as treatment references.

• Reproductive and Developmental Biology
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• v.36 no.4
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• pp.269-273
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• 2012

The present study was to assess the in vitro viability and sexing rate of bovine embryos. Blastocysts were harvested on day 7~9 day after insemination(in vitro and in vivo), and the sex of the embryos was examined using the LAMP method. Embryo cell biopsy was carried out in a $80{\mu}l$ drop $Ca^{2+}$, $Mg^{2+}$ free D-PBS and, biopsied embryos viability were evaluated after more 12 h culture in IVMD culture medium. The formation of recovered embryo to expanded and hatching stages had ensued in higher of sexed embryo in vivo than in vitro (100% vs. 89%, p<0.05), and in vitro, the rates of degeneration after sexing were significantly (p<0.05) higher in vitro than in vivo(11% vs. 0.0%). The rates of the predicted sex were female 61% vs. 56%, and male 39% vs. 44% in vivo and in vitro, respectively. The rates of survival following different biopsy methods were seen between punching and bisection method in vivo and in vitro (100% vs. 89% and 100% vs, 78% respectively). Biopsy method by punching was significantly (p<0.05) higher than bisection between produced embryos in vivo and in vitro. The present data indicate that with microblade after punching for embryo sexing results in high incidence of survivability on development after embryo biopsy. It is also suggested that LAMP-based embryo sexing suitable for field applications.

• Journal of Embryo Transfer
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• v.22 no.3
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• pp.191-194
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• 2007

The purpose of the present study was to set up basic information of size and status of ovaries by using ultrasonography to retrieve in vivo matured oocytes with ovum pick- up method. Ovaries were collected from the abattoir in Jeju in May and June which is breeding season. When the size of ovaries on ultrasonography was compared with real size measured by caliper, no significant difference was shown (p<0.05). The number of preovulatory follicles (>21mm) was investigated with ultrasonography and naked eyes. Ultrasonography group had 0.83 preovulatory follicles per ovary and naked-eye group had 0.75 preovulatory follicle per ovary and their average size was 2.86 cm and 2.34 cm, respectively. The average number of follicle was 4.25 with ultrasonography and 4.38 with naked eyes. There was no significant difference considering the size of follicle and number of follicle between ultrasonography and actual size except for the size of preovulatory follicle, suggesting that information of ultrasonography is able to use for OPU or other reproductive technology of mare.

• Korean Journal of Poultry Science
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• v.39 no.1
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• pp.9-15
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• 2012

The aim of this study was to investigate phenotypic characteristics, morphometric measurements, reproduction and production performances of Aseel chicken of Bangladesh. The dominant feather color of neck/hackles was red in both males (56.14%) and females (54.16%) while the sickle feather color was mostly black in both chickens (71.93% vs. 54.17%). The predominant saddle and breast feather colors were red (40.35%) and black (64.91%), respectively, in male whereas most frequent observed color was pale brown in female (58.33 and 50.0%, respectively). The predominant feather color of wing bow and wing bay was found black (68.42 and 80.70%, respectively) in male but only pale brown color was observed in females (62.5 and 54.17%, respectively) for these two characters. Different phenotypic measurements such as the average shank length and circumference were $12.79{\pm}0.13$ and $7.8{\pm}0.08$ cm, respectively, in male and $10.21{\pm}0.25$ and $5.81{\pm}0.21$ cm, respectively, in female. Keel length was $14.39{\pm}0.19$ cm in male and $10.79{\pm}0.23$ cm in female. The average adult live weight in male was measured $3749.12{\pm}83.44$ g while in female it was $2062.50{\pm}105.26$ g. The age of 1st lay was found to be 28.86 weeks. Total number of eggs laid per year ranged between 24~48, number of clutch/hen/year varied from 2 to 4 and number of eggs/clutch/hen was found to be 10~12. The average live weight of Aseel chicken at 1, 2, 3, 4, 6, 8, 10, 12, 16 and 17 weeks of age were recorded as $31.14{\pm}0.55$, $48.63{\pm}3.99$, $116.57{\pm}5.72$, $138.40{\pm}5.91$, $212.88{\pm}4.82$, $361.00{\pm}9.72$, $577.50{\pm}42.86$, $743.75{\pm}24.65$, $1086.00{\pm}26.02$, $1402.00{\pm}24.54$ and $1432.00{\pm}27.00$ g respectively. Finally, this phenotypic characterization as well as productive and reproductive performances of Aseel chicken will give the baseline information to researcher for further study and for planning any on-ward conservation and implement strategy.

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.181-186
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• 2005

This experiment was conducted to investigate effects of the two different in vitro production systems, serumcontaining system (IVM, IVF and IVC; TCM199, TALP and CR1aa) and serum-free system (IVM, IVF and IVC; IVMD101, IVMD100 and IVMD101), on the development of in vitro fertilized or DNA-microiniected embryos. We also examined the effect of DNA dosage and its expression pattern in embryos. The DNA used for microinjection was a green fluorescence protein gene. The development rates to $\geq$ 2cell, 8cell and blastocyst stage were significantly higher in vitro fertilized embryos than those in DNA-microinjected embryos. The development rate to the 8-cell stage was significantly higher in serum-free system than in serum-containing system (p<0.05; $3.3\%\;vs.\;15.5\%\;and\;21.4\%$, respectively). The development rates to the blastocyst stage of in vitro fertilzed or DNA-microinjected embryos between two different culture system ($2.7\%\;vs.\;2.3\%\;and\;23.0\%\;vs.\;23.6\%$, respectively) were not different. The development rates of embryos injected 2 ng/uL DNA was higher. than those of embryos injected 4 or 8 ng/uL DNA. The GFP expression rate of 1-cell embryos was significantly higher than that of 2-cell and 4-cell embryos, whereas the rates were not different between 4-cell and blastocyst-stage embryos.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.281-285
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• 2011

In the present study, we identified differentially methylated region (DMR) upstream of Dnmt1o and Dnmt1s gene in early porcine embryos. Porcine Dnmt1o had at least one DMR which was located between -530 bp to -30 bp upstream from transcription start site of the Dnmt1o gene. DNA methylation analyses of Dnmt1o revealed the DMR to be hypomethylated in oocytes, whereas it was highly methylated in sperm. Moreover, the DMR upstream of Dnmt1o was gradually hypermethylated from oocytes to two cells and dramatically changed in the methylation pattern from four cells to BL stages in an in vivo. In an IVF, the methylation status in the DMR upstream of Dnmt1o was hypermethylated from one cell to eight cells, but demethylated at the Morula and BL stages, indicating that the DNA methylation pattern in the Dnmt1o upstream ultimately changed from stage to stage before the implantation. Next, to elucidate whether DNA methylation status of Dnmt1s upstream is stage-by-stage changed in during porcine early development, we analyzed the dynamics of the DNA methylation status of the Dnmt1s locus in germ cell, or one cell to BL cells. The Dnmt1s upstream was highly methylated in one and eight cells, while less methylated in two, four, morula, and BL cells. Taken together, our data demonstrated that DNA methylation and demethylation events in upstream of Dnmt1o/Dnmt1s during early porcine embryos dramatically occurred, and this change may contribute to the maintenance of genomewide DNA methylation in early embryonic development.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.197-201
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• 2011

This study was conducted to investigate the effects of donor cell treatments on the production of transgenic cloned piglets. Ear fibroblast cell obtained from NIH MHC Inbred minipig was used as control. The GalT knock-out/CD45 knock-in (GalT/CD46) transgenic cell lines were established and used as donor cells. The reconstructed GalT/CD46 embryos were surgically transferred into oviduct of naturally cycling surrogate sows (Landrace ${\times}$ Yorkshire) on the second day of standing estrus. Unlike control (1.2 kV/cm, 75.4%), the fusion rate of the GalT/CDl6 donor cells was significantly higher in 1.5 kV/cm, (84.5%) than that of 1.25 kV/cm, (20.2%) (p<0.01). When the number of the transferred embryos were more than 129, the pregnancy and delivery rates were increased to 13/20 (65%) and 5/20 (25%) compared to less then 100 group [1/6 (16.7%) and 0/6 (0%)], respectively. To analyze the effect of donor cell culture condition on pregnancy and delivery rates, the GalT/CD46 donor cells were cultured with DMEM or serum reduced medium. In serum reduced medium group, the pregnancy and delivery rates were improved to 8/12 (66.7%) and 5/12 (41.7%) compared to DMEM group [3/7 (42.9%) and 0/7 (0%)], respectively. In conclusion, it can be postulated that an appropriate fusion condition and culture system is essential factors to increase the efficiency of the production of transgenic cloned piglets.

• Reproductive and Developmental Biology
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• v.38 no.3
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• pp.129-136
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• 2014

This study was carried out to elucidate the effect of antibiotic-free rearing system(ARS) on cortisol level, stress hormone, and fatty acid content in the edible muscle tissues, that were of M. longissimus in Hanwoo. These cattle were reared in two different systems including antibiotic-free (ARS) and conservative system (CRS). To increase the experimental reliability, the muscle samples were purchased 3 times from 3 Korean brands of beef produced with ARS or CRS. In the muscle tissue, cortisol level was significantly lower in ARS than CRS, (p=0.0176). But the levels of total saturated- and unsaturated-fatty acids were not significantly different between ARS and CRS (p>0.05). Of total fatty acids, the total saturated fatty acid tended to be greater in CRS and the total unsaturated fatty acid tended to be greater in ARS. However, of the total unsaturated fatty acids, the level of n-6 unsaturated fatty acids was significantly higher in ARS than CRS (p=0.0040). Especially, ${\alpha}$-linolenic acid (ALA) and ${\gamma}$-linolenic acid (GLA) levels were significantly higher in ARS (p<0.01). The n-6 fatty acid content and cortisol level in muscle tissue were negatively correlated at p=0.0140. In conclusion, ARS may produce beef with higher quality which contains lower cortisol and greater n-6 fatty acids, such as ALA and GLA.

• Journal of Animal Science and Technology
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• v.49 no.1
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• pp.15-24
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• 2007

The purpose of this study was to evaluate the presence and differential expression of connexin isoforms in the efferent ductules (ED) of male rat reproductive tract during postnatal development. Total RNA was isolated from the ED collected from rats at 1 week, 2 weeks, 1 month, 3 months, and 6 months of ages. Expression of six connexin mRNAs of 14 isoforms tested was detected by semi-quantitative RT-PCR. Fluctuation of mRNA levels of connexins 26 and 30 was found according to ages. A significant decrease of connexin 31.1 mRNA level was observed in the ED at 1 month of age. The highest levels of connexin 37 and 45 mRNAs were detected in the ED of early developmental period, while the expression of connexin 43 was the lowest at 1 week of age. The present study demonstrates differential regulation on expression of connexin isoforms in the rat ED in age-dependent manner.

• Reproductive and Developmental Biology
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• v.30 no.2
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• pp.135-141
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• 2006

The present study was conducted to investigate the effects of cumulus cells and porcine follicular fluid (pFF) on plasminogen activator (PA) activity and oocytes maturation in vitro in the pig. The cumulus-oocyte complexes (COCs) and denuded oocytes (DOs) were incubated in NCSU-23 medium with or without 10% pFF for 0, 24, or 48 hr. In the presence of cumulus cells, the proportions of oocytes matured to metaphase-II stage were significantly (P<0.05) higher in medium with pFF than without pFF (69.8 vs. 37.7%, respectively). When COCs and DOs were cultured in the presence of pFF, tissue-type PA (tPA), urokinase-type PA (uPA), and tPA-PA inhibitor (tPA-PAI) were observed in COCs, and PA activities were higher at 48 hr than 24 hr. When COCs and DOs were cultured in the absence of pFF, tPA and tPA-PAI were observed in COCs, and PA activities were increased as duration of culture increased. No PA activities were detected in DOs regardless of pFF supplementation. When porcine oocytes were cultured in the presence of pFF for 24 and 48 hrs, the activities of tPA-PAI, tPA, and uPA were observed in both COCs and DOs. In medium of absence of pFF, PA activities were observed in oocytes with cumulus cells only. On the other hand, three plasminogen-dependent lytic bands (tPA-PAI, tPA, and uPA) were observed in pFF cultures. Particularly uPA activity was higher than the other kinds of PA activity. When oocytes and cumulus cells were separated from porcine COCs at 0 hr of culture, tPA-PAI, tPA, and uPA were detected in cumulus cells at 48 hr of culture, but no PA activities were in DOs. The presence of pFF and cumulus cells in maturation medium stimulated not only nuclear and cytoplasmic maturation in porcine COCs, but also PA production by cumulus cells and COCs. It is possible that PAs produced by cumulus cells migrated through the gap junction between oocyte and cumulus cells. These results suggest that porcine oocytes have no ability to produce PA themselves.