• 한국가축번식학회지
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• 제21권1호
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• pp.61-69
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• 1997

본 연구는 EGF와 anti-EGF가 초기 생쥐배아의 발생 및 부화에 미치는 영향을 알아보고자 실행되었다. 초기 2세포기부터 상실배까지의 배아를 EGF와 anti-EGF를 각각 처리한 Ham's F10 배양액에서 배양하여 그 발생률과 부화율을 대조군과 비교하였다. EGF 처리시 배양시간에 따른 발생률은 증진되었으나 통계학적 유의성은 없었다. EGF 처리군에서의 부화율(57.5, 62.5, 65.0, 62.5%)은 대조군(35%)에 비하여 유의하게 (p<0.01) 높았다. Anti-EGF 처리시 각 발생시기별 1:1000 실험군의 발생률은 대조군과 차이가 없었다. 그러나, 1:100 실험군의 경우, 2∼4세포기의 배아는 모두 4∼8세포기에서 정지되었고, 8세포기와 상실배의 포배형성은 48시간 이상 지연되었으며 부화 역시 대조군에 비해 억제되었다(8세포기; 2%, 44%, 상실배; 6.2%, 58.3%). 이 실험에서, EGF는 생쥐 배아의 포배형성과 부화를 증진시키는 반면, anti-EGF는 이를 억제하였다. Anti-EGF 처리시 나타나는 발생정지 현상은 anti-EGF가 배아에서 만들어지는 EGF와 반응하여 EGF의 작용을 억제시키기 때문으로 사료된다. 그러므로 EGF는 paracrine mode로서 뿐만 아니라 antocrine mode로서 생쥐 초기배아의 발생에 중요한 인자로 작용함을 알 수 있었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제5권1호
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• pp.1-12
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• 2002

Ecophysiologically diapause represents a syndrome of physiological and biochemical characteristics, all of which ensure survival during a long period of dormancy. Since, silkworm enters diapause as embryo at the early embryonic stage, the duration of egg life depends on the duration of embryonic diapause. The nature of diapause in silkworm, Bombyx mori, is primarily determined by genetic characters and endocrinologicnl mechanisms, mediated by environmental factors such as temperature and photoperiod. Hibernating potency value besides nucleic acid and carbohydrate metabolism, production and utilization of sorbitol are also equally responsible for induction, initiation, determination, maintenance and termination of diapause. Embryonic diapause in Bombyx moir, induced by active secretion of sub-oesophageal ganglion is attributed to hormonal system and metabolic adjustment, which serves to bring about a new physiological state. Metabolic conversion of trehalose to glycogen at induction, glycogen to sorbitol at initiation and sorbitol to glycogen at termination of diapause is correlated and in each metabolic shift a key enzyme becomes active in response to hormonal and environmental stimulation. An attempt has been made in this review article to discuss briefly the nature of embryonic diapause, influence of various factors on diapause nature, hormonal mechanism of diapause besides biochemical composition of egg, nucleic acid and carbohydrate metabolism, production and utilization of sorbitol in relation to induction, determination, maintenance, initiation and termination of diapause in the silkworm, Bombyx mori.

• Plant Biotechnology Reports
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• 제3권4호
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• pp.341-345
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• 2009

This study describes culture conditions for a plant regeneration system via a combined pathway of somatic embryogenesis and organogenesis in root explant cultures of the commercial rose cultivar 'Charming'. Root explants formed white calluses at a frequency of 30% after 6 weeks of culture on Schenk and Hildebrandt (SH) medium supplemented with $11mg\;1^{-1}$ 2,4-dichlorophenoxyacetic acid. After 6 weeks of transfer to SH medium without growth regulators, initial white calluses gave rise to globular somatic embryos at a frequency of 2.8%, which were subsequently dedifferentiated to embryonic tissues. Somatic embryos or embryonic tissues initially derived from root explants did not undergo development beyond cotyledonary stage. To produce adventitious shoots, embryonic tissues were sliced and cultured on SH medium with $0.5mg\;1^{-1}$ 6-benzyladenine. After 4 weeks of culture, 28% of embryonic tissue explants formed adventitious shoots. Regenerated shoots were rooted on half strength SH medium with $0.1mg\;1^{-1}$ ${\alpha}-naphthalaneacetic$ acid and subsequently grown to maturity. Root-derived embryonic tissues were proliferated by subculture, while retaining the capacity for shoot production for a few years.

• 한국수정란이식학회지
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• 제27권2호
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• pp.121-126
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• 2012

The objective of this study was to examine the effect of in vitro culture media on embryonic development of in vitro-matured (IVM) oocytes after parthenogenetic activation (PA) in pigs. Immature pig oocytes were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones for the first 22 h and then further cultured in hormone-free medium for an additional 22~26 h. IVM oocytes were activated by electric pulses and cultured in porcine zygote medium-3 (PZM-3) and North Carolina State University-23 supplemented with essential and non-essential amino acids (NCSU-23aa). These media were further modified by supplementing 2.77 mM myo-inositol, 0.34 mM trisodium citrate, and $10{\mu}M$ ${\beta}$-mercaptoethanol (designated as mPZM-3 and mNCSU-23aa, respectively). Culture of PA embryos in mPZM-3 significantly increased development to the blastocyst stage than culture in NCSU-23aa (36.2% vs. 24.8%, p<0.05). Modified PZM-3 showed a significantly higher blastocyst formation than NCSU-23aa in both groups of embryos that were activated at 44 h and 48 h of IVM (51.0% vs. 35.5% and 49.0% vs. 34.2% in oocytes activated at 44 h and 48 h of IVM, respectively). Irrespective of the follicle diameter where oocytes were collected, embryonic development to the blastocyst stage was increased (p<0.05) by the culture in mPZM-3 compared to culture in NCSU-23aa (25.9% vs. 34.2% and 32.9% vs. 44.8% in embryos derived from small and medium size follicles, respectively). Our results demonstrated that culture media had significant effect on preimplantation development PA embryos and that mPZM-3 was superior to mNCSU-23 in supporting development to the blastocyst stage in pigs. This beneficial effect of mPZM-3 on embryonic development was not impaired by other factors such as time of oocyte activation and origin of immature oocytes (small and medium size follicles).

• 한국가축번식학회지
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• 제26권4호
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• pp.385-394
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• 2002

This study was constructed the correlations of the embryonic developmental rates and the frequency of chromosome aberration using ear-skin-fibroblast cell in nuclear transfer (NT) derived embryos. Karyoplast-oocyte complexes were fused and activated simultaneously, then cultured for seven days to assess development. The developmental rates of NT and in vitro fertilization (IVF) embryos were 55.4% vs 63.5%, 31.7% vs 33% and 13.4% vs 16.8% in 2 cell, 8 cell and blastocyst, respectively. Firstly, the frequency of chromosome aberrations were evaluated using fluorescent in situ hybridization (FISH) technique with porcine chromosome 1 submetacentric specific probe. Chromosome aberration was detected at day 3 on the embryo culture, the percentages of chromosomal aneuploidy in NT and IVF embryos at 4-cell stage were 40%, 31.3%, respectively. Secondly, embryonic fragmentation was evaluated at 4-cell stage embryo. Frequency of embryonic fragmentations was in 51.3% of NT, 61.3% of IVF, 28.9% of parthenogenetic activation at 4-cell stage. The proportion of fragmentation in NT embryos was higher than activation embryos. This result indicates that chromosomal abnormalities and embryonic fragments are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT related with lower implantation rate, increased abortion rate and production of abnormal fetuses.

• Animal cells and systems
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• 제13권4호
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• pp.411-418
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• 2009

We are reporting the identification, expression patterns, and possible biological functions of zebrafish kif3b (kif3bz) encoding 475 amino acids. Kif3Bz contains the kinesin motor domain, catalytic domain, KISc domain, and one single coiled coil domain. Phylogenetic analysis indicates that kif3bz is a highly conserved gene among the tested vertebrates. First of all, both maternal and zygotic messages of kif3bz were evenly distributed in the blastomeres at 2-cell stage. Its ubiquitous expression throughout the blastomeres continued till 40% epiboly. However, kif3bz transcripts became restricted in Kupffer's vesicle at tailbud and 6-somite stages. At 13-somite stage, kif3bz expression pattern became specific to the telencephalon, diencephalon, trigeminal placode, and somites. Such expression patterns were further intensified in the telencephalon, diencephalons, hind brain, pronephric ducts, optic vesicles, and spinal cord neurons in the 23-somite stage embryos, and last till 24 hpf. We discussed possible functions of Kif3Bz related to the vertebrate embryonic development.

• Asian-Australasian Journal of Animal Sciences
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• 제23권7호
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• pp.873-879
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• 2010

We investigated the effect of various concentrations of taurine during in vitro fertilization (IVF) on the embryonic development up to the blastocyst stage of bovine oocytes fertilized with three different Japanese Black bulls (Bull A, B and C). In vitro matured oocytes were fertilized with various concentrations of taurine (0, 1, 10, 50 and 100 mM) in the presence of 2.5 or 5.0 mM caffeine plus $25{\mu}g$/ml heparin (CH) for 6 hr or $100{\mu}g$/ml heparin (H) for $24{\pm}2$ h. After IVF, the cleavage rates from the 2 to 16 cell stage determined at 3 days and the development rates up to the blastocyst stage determined at 7-8 days from the onset of IVF were assessed. Although the cleavage rates for the taurine concentration groups were not significantly increased in any of the three bulls in the CH groups, the development rates up to the blastocyst stage of the 50 mM taurine group of Bulls A and B, and of the 1 to 50 mM groups of Bull C were increased (p<0.05) compared to those of the control (0 mM taurine) groups. On the other hand, none of the bulls in the H groups showed any significant increase either in the cleavage rates or blastocyst formation rates in any taurine concentrations groups compared with those of the control groups. These results indicate that the addition of 50 mM taurine to a fertilization medium containing caffeine and heparin may stimulate embryonic development up to the blastocyst stage when fertilized with different bull semen.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.276-276
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• 2004

Pluripotent stem cells have been generated from two embryonic sources. ES cells are generated from ICM of blastocyst stage embryos, and embryonic germ (EG) cells are generated from primordial germ cells (PGCs). Both ES and EG cells are pluripotent and present important characteristics such as high levels of alkaline phosphatase (AP) activity, multi-cellular colony formation, normal and stable karyotypes, continuously passaging ability, and the capability of differentiation into all three embryonic germ layers. (omitted)

• Journal of the Korean Data and Information Science Society
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• 제16권4호
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• pp.951-958
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• 2005

As a foundation study of stem cell applied research, it is necessary to identify the large gene expression through cDNA microarray to understand principles of the level of molecular about cell function. In this paper, we investigated the gene expression through the K-means clustering method and path analysis with genes related to pluripoteny and differentiation in an mouse early stage embryonic development process and embryonic stem cell differentiation. We find a few biological phenomenon through this study. Also, we realize that this process provides functional relationship of unknown genes.

• Clinical and Experimental Reproductive Medicine
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• 제32권2호
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• pp.133-147
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• 2005

Objectives: This study was carried out to establish human embryonic stem cells derived from frozen-thawed embryos using mouse embryonic fibroblasts (mEFs), human fetal skin and muscle fibroblasts as feeder cells, and to identify the characteristic of embryonic stem cells. Methods: When primary mEFs, human fetal skin and muscle fibroblasts were prepared, passaging on 4 days from replating could have effective trypsinization and clear feeder layers. Eight of 23 frozenthawed 4~8 cell stage embryos donated from consenting couples developed to blastocysts. Inner cell mass (ICM) was isolated by immunosurgery. ICM was co-cultured on mEFs, human fetal skin or muscle fibroblasts. The ICM colonies grown on mEFs, human fetal skin or muscle fibroblasts were tested the expression of stage specific embryonic antigen-3, -4 (SSEA-3, -4), octamer binding transcription factor-4 mRNA (Oct-4) and alkaline phosphatase surface marker. Results: We obtained 1 ICM colony from 2 ICM co-cultured on mEFs as feeder cells and did not obtain any ICM colony from 6 ICM clumps co-cultured on human fetal skin or muscle fibroblasts. The colony formed on mEFs could be passaged 30 times every 5 days with sustaining undifferentiated colony appearance. When the colonies cultured on mEFs were grown on human fetal skin or muscle fibroblasts, the colonies could be passaged 15 times every 9 days with sustaining undifferentiated colony appearance. The colonies grown on mEFs and human fetal fibroblasts expressed SSEA-4 and alkaline phosphatase surface markers and positive for the expression of Oct-4 by reverse transcription-polymerase chain reaction (RT-PCR). The produced embryoid body differentiated spontaneously to neural progenitorlike cells, neuron-like cells and beating cardiomyocyte-like cells, and frozen-thawed embryonic stem cells displayed normal 46,XX karyotype. Conclusions: The human embryonic stem cells can be established by using mEFs and human fetal fibroblasts produced in laboratory as feeder cells.