• Korean Journal of Animal Reproduction
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• v.17 no.3
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• pp.233-242
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• 1993

The effects of in vitro maturation and sperm treatment condition on the in vitro fertilization (IVF) and developmental capacity of bovine oocytes were investigated and the development of embryos was compared under the 2 different co-culture system, with GC or BOEC. The cultured embryo to 16 cell or morula wre transferred into recipients or frozen by 2 different freezing method. The results obtained were summarized as follows; 1. In vitro maturation rates of vovine follicular oocytes cultrued in TCM199 with 10% FCS or ECS were 64.0% and 72.7%, but the case of addition of 10% FCS or ECS to TCM199 co-cultured with granulosa cells were 81.3% and 84.0%, respectively. IVM rate of three TCM199 added to granulosa cells was higher than that of media without granulosa cells. 2. When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC and then fertilized in vitro by sperm treated with caffeine, embryo developments of bovine oocytes co-cultured with BOEC were 38.4% and 51.4%, respectively. But those of bovine oocytes co-cultured with GC were 52.2% by sperm treated with caffeine-heparin. 3. Cleavage rates of bovine oocytes cultured with 10% FCS alone and fertilized in vitro by sperm treated with caffeine-heparin was 33.0%. 4. When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC, embryo developments of bovine ooctyes co-cultured with BOEC of GC were 46.0% and 50.2%, respectively. 5. When bovine follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments co-cultured with BOEC or GC were 45.2% and 51.4%, respectively. 6. When Korean Native cow's follicular oocytes matured in TCM199 with 10% FCS and GC, embryo developed co-cultured with BOEC or GC were 45.2% and 51.4%, respectively. 6. When Korean Native cow's follicular oocytes matured in TCM199 with 10% FCS and GC, embryo developments of the bovine oocyte co-cultured with BOEC and GC were 41.8% and 60.1%. But with FCS 10% those of the bovine oocytes co-cultured with BOEC and GC were 42.0% and 48.4%, respectively. 7. When Holstein's follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments fo the bovine oocytes co-cultured with BOEC and GC were 50.0% and 57.7%, but with ECS 10% those of the bovine oocytes co-cultured with BOEC and GC were 52.2% and 56.5%, respectively. 8. The viability of frozen-thawed embryos ranged from 60~80% and those of frozen-thawed embryos from vitrification was lower than that from conventional metiod. 9. The selected fresh embryos were transferred nonsurgically to 7 recipients but did not result in pregnancy.

• Korean Journal of Animal Reproduction
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• v.20 no.3
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• pp.333-341
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• 1996

One of the biggest problems involved in transgenic animal production is lack of appropriate market genes. To overcome this problem, we tested whether the genes of SEAP (secreted alkaline phosphatase) and GFP (green fluorescence protein) on our retrovirus vectors can be applicable to the transgenic animal production. The main advantage of these marker genes over other generally mainpulation can be selected without sacrificing viability. The results obtained in this study are summarized as follows: 1. Removal of zona pellucida from the mouse zygotes did not affect embryo developments to blastocysts. 2. Co-culture of zona-free embryos with virus-producing cells for 6 hours also did not affect embryo developments to blastocysts. 3. Among 58 blastocysts developed from the zona-free zygotes co-cultured with the virus-producing cells, SEAP expression was observed from the 6 blastocysts. 4. Expression of the GFP gene was detected from the virus- producing cells but no embryo expressing the gene was counted among 50 blastocysts developed from the zona-free zygotes co-cultured with the virus-producing cells.

• Reproductive and Developmental Biology
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• v.41 no.2
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• pp.33-40
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• 2017

X-chromosome inactivation is one of the most complex events observed in early embryo developments. The epigenetic changes occurred in female X-chromosome is essential to compensate dosages of X-linked genes between males and females. Because of the relevance of the epigenetic process to the normal embryo developments and stem cell studies, X-chromosome inactivation has been focused intensively for last 10 years. Initiation and regulation of the process is managed by diverse factors. Especially, proteins and non-coding RNAs encoded in X-chromosome inactivation center, and a couple of transcription factors have been reported to regulate the event. In this review, we introduce the reported factors, and how they regulate epigenetic inactivation of X-chromosomes.

• Journal of Environmental Health Sciences
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• v.31 no.4 s.85
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• pp.254-259
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• 2005

Permethrin, a synthetic pyrethroid insecticide, has been widely used to protect domestic animals and the public health, as well as in agriculture against a variety of pests, which provides potential for environmental exposure. Permethrin is classified as possible human carcinogen and endocrine disrupting chemical by many international authorities. However, its developmental effects have been rarely studied. This study investigated the effects of permethrin during embryo-genesis. Developmental toxicity of permethrin was evaluated using short-term in vitro battery system. Gestation day 9.5 rat embryos (organogenesis) were cultured with permethrin (0.1,0.4 and 0.8 mg/ml) for 48 hours using whole embryo culture system. All the treatments exhibited significant decreases in the total morphological score. Permethrin induced significant growth retardation and the developmental abnormality at doses of 0.4 and 0.8 mg/ml. Moreover, the DNA and protein contents of embryos decreased in dose-dependent manner. These observations suggest that permethrin contributes to toxicity on embryonic developments in rats.

• Journal of Ginseng Research
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• v.16 no.1
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• pp.37-43
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• 1992

Structural changes of the endosperm of Panax ginseng C.A. Meyer from fertilization to germination were investigated by light microscope. The endosperm of the ginseng seed is cellular type. Since endosperm cells adjacent embryo continuously breakdown and disappear with the elongation of embryo, the real of endosperm is gradually decreased. As the anatropous ovules of immature seed with green seed coat developes more and more, ovary cells adjacent ovary cavity become abundant by the periclinal division, their size is decreased, hypotrophy of cell wall discern, and they are gradually differentiated in seed coat. Though embryo responds strongly to basic dye at the stage of completion of endosperm formation, tissue of endosperm responds to acidic dye positively Cell wall of embryo and endosperm are composed of primary cell wall not lignified. Endosperm cells adjacent embryo begin to breakdown in the endosperm tissue of indehiscent seed before the beginning of the after-ripening. Dehiscent seed of which seed coat is opened through after-ripening represent the form as a seedling in the result of embryo developments with the formation of organs; radicle, cotyledon, plumule. Umbilifom layer represents strong positive response to the toluidine blue and the basic function. Umbiliform layer that endosperm cells breakdown and disappear is observed clearly at the periphery of the embryo cotylemon, while slightly at the periphery of the radicle.

• Biomedical Science Letters
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• v.10 no.3
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• pp.305-308
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• 2004

We have examined alcohol-induced malformation of chick embryo. Alcohol was considered to induce the malformation of developing embryo and to have bad effects on embryonic stage. We injected alcohol into air sac on day 4 of incubation. Ten ％ alcohol-treated group showed a little decrease on their body length compared to the untreated group and distilled water-treated group. Thirty ％ alcohol-treated group showed significant decrease on their body length compared to the untreated group and 10％ ethanol treated group. In addition, we have observed malformation of eyeballs and bills. These results indicate that alcohol affects chicken developments and brings on malformation of developing stage.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.63-68
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• 1994

To confirm the overcome of in vitro 2-cell block, ICR mouse I-cell embryos were cultured in CZB media. All embryos in CZB were overcome in vitro 2-cell block and 92% of embryos were developed to the blastocyst at day 4. However, in m-KRB group(control) only 20% of embryos were developed over 2-cell. Any embryos in m-KRB did not develop to the morular stage. Developments and degenerations of ICR mouse I-cell embryos were compared in CZB medium prepared with water of three quality:(l) Milli-Q ultrafiltration water(UF);(2) Milli-Q reverse osmosis water(RO);(3) tap water(TAP). The objective was to evaluate the potential of quality control using ICR mouse 1-cell embryos. The more water was purified, the better embryo developments were supported and the less embryos were degenerated. As a quality control system, the culture of ICR 1-cell mouse embryos in CZB was useful.

• Journal of Embryo Transfer
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• v.25 no.3
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• pp.189-193
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• 2010

We investigated the cleavage rate and blastocyst yield for each culture condition to enhance tolerance of cryo-preservation of bovine IVF embryo with relatively lower cryo-tolerance compared to in vivo embryo. The cleavage rate and blastocysts yield for CR1aa, IVMD, IVD, CR1aa+10% FBS were 73.2, 69.3, 72.8, 68.5% and 44.1, 30.8, 33.3, 48.0%, respectively. The values did not differ among each treatments without serum. For embryo vitrification, In vivo and In vitro blastocysts were exposed to VS1(10% glycerin, 0.1 M glucose, 0.1 M sucrose, PEG 1%) for 5 min, and VS2 (10% glycerin, 10% EG, 0.2 M glucose, 0.2 M sucrose, PEG 2%) for 5 min and then VS3 (10% glycerin, 30% EG, 0.3 M glucose, 0.3 M sucrose, PEG 3%) for 1 min. The exposed embryos were then loaded into the 0.25 ml plastic straws and then plunged into liquid nitrogen. The straws were held for period of 1 to 2 weeks before thawing. In embryo viability, no differences in blastocyst re-expansion rates were found between in vivo and in vitro embryos. whereas expansion-BL rates was significantly higher for in vivo-derived embryos (72.7%) when compared to in vitro-derived embryos (51.4%), respectively (P<0.05). In conclusion, our results indicate that combined use of CRIaa culture medium with vitrification might enhance tolerance of cryopreservation for bovine IVF embryo production.

• Journal of Embryo Transfer
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• v.29 no.2
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• pp.141-148
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• 2014

Quercetin and genistein, plentifully present in fruits and vegetables, are flavonoid family members that have antioxidative function and plant-derived phytoestrogen activity. The antioxidative effects of quercetin and genistein on boar sperm characteristics and in vitro development of IVF embryo were investigated. The sperm motility was increased by addition of genistein $50{\mu}M$ for 6 hr incubation compared to control (p<0.05). The sperm viability was increased by addition of quercetin 1 and $50{\mu}M$ and genestein 1 and $50{\mu}M$ for 3 hr incubation. In addition, the sperm viability seemed to be increased dose-dependantly by addition of quercetin or genistein 1 and $50{\mu}M$, respectively (p<0.05). The membrane integrities were not increased by quercetin or genistein treatments for 3 hr or 6 hr incubation period except for quercetin $1{\mu}M$ for 3 hr incubation. In mitochondrial activities, addition of quercetin $50{\mu}M$ for 6 hr incubation increased mitochondrial activity but decreased at $100{\mu}M$ concentration compared with control (p<0.05). When porcine IVF embryos were cultured in PZM-3 medium supplemented with low concentrations of quercetin ($1{\sim}10{\mu}M$), the developmental rates to morula and blastocyst increased but significantly decreased at high concentrations of quercetin ($25{\sim}50{\mu}M$). The highest developmental rate to blastocysts among all concentrations of quercetin was shown at quercetin $10{\mu}M$ (p<0.05). The developmental rates to morula or blastocysts at low ($0.01{\sim}1{\mu}M$) and high ($5{\sim}10{\mu}M$) concentrations of genistein were not significantly different among all treatment group and genistein did not affect on IVF embryo development. These results suggest that quercetin and genistein seem to have positive effects at certain concentrations on sperm characteristics such as motility, viability and mitochondrial activity. In addition, low concentrations of quercetin (1, 5 and $10{\mu}M$) in this experiment, seem to have beneficial effect on porcine IVF embryo development but genistein did not affect on it at all given concentrations ($0.01{\sim}10{\mu}M$).

• Korean Journal of Plant Tissue Culture
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• v.26 no.4
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• pp.275-280
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• 1999

In order to find optimum conditions for somatic embryogenesis from different individual (2-year-old) in Aralia elata were cultured on MS medium supplemented with 1.0 mg/L 2,4-D, 3% sucrose, and 0.3% gelrite. We also investigated the effect of MS medium salt concentration, BA and ABA on the embryo germination and plant regeneration. While noticeable difference was observed on somatic embryo induction among different individual tree, no apparent difference was seen in both germination and regeneration frequencies. Compared with nonembryogenic calli, embryogenic calli tended to look yellow and/or pale brown in color, slowly growing and soft in their texture. Regardless of BA or ABA treatment, half-strength MS salt medium proved to be better than full strength MS medium in both embryo germination and plant regeneration. Both hypocotyl and cotyledon developments were slightly promoted by adding 0.1 mg/L BA. However, its effect on germination and regeneration seemed inferior to control. ABA treatment on somatic embryos at their torpedo and early cotyledonary stages resulted in poor response in germination and regeneration. Although most regenerated plantlets varied greatly in cotyledon number and shape, they could be developed into normal plants after 4 weeks in culture. More than 95% plantlets were acclimatized in an artificial soil mixture, successfully transplanted to nursery bed and grew normally without any phenotypic abnormalty.