• Title/Summary/Keyword: co-culture system

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Effect of a short-term in vitro exposure time on the production of in vitro produced piglets

  • Hwang, In-Sul;Kwon, Dae-Jin;Kwak, Tae-Uk;Lee, Joo-Young;Hyung, Nam-Woong;Yang, Hyeon;Oh, Keon Bong;Ock, Sun-A;Park, Eung-Woo;Im, Gi-Sun;Hwang, Seongsoo
    • Journal of Embryo Transfer
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    • v.31 no.2
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    • pp.117-121
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    • 2016
  • Although piglets have been delivered by embryo transfer (ET) with in vitro produced (IVP) embryos and blastocysts, a success rate has still remained lower level. Unlike mouse, human, and bovine, it is difficult to a production of piglets by in vitro fertilization (IVF) because of an inappropriate in vitro culture (IVC) system in pig. Therefore, the present study was conducted to investigate whether minimized exposure time in IVC can improve the pregnancy and delivery rates of piglets. Immediately after IVM, the oocytes were denuded and co-incubated with freshly ejaculated boar semen for 3.5 to 4 hours at $38.5^{\circ}C$ under 5% $CO_2$ in air. To avoid long-term exposure to in vitro state, we emitted IVC step after IVF. After that the presumptive zygotes were transferred into both oviducts of the surrogate on the same day or 1 day after the onset of estrus. Pregnancy was diagnosed on day 28 after ET and then was checked regularly every month by ultrasound examination. The 3 out of 4 surrogates were determined as pregnant (75%) and a total of 5 piglets (2 females and 3 males) were delivered at $118.3{\pm}2.5$ days of pregnancy period. In conclusion, a short-term exposure time may be an important factor in the production of IVP-derived piglets. It can be apply to the in vitro production system of transgenic pig by IVF, cloning, and pronuclear microinjection methods.

Studies on the Improvement of Performance and Reproductive Efficiency in Farm Animal Ⅵ. Studies on Improvement of Development Potential of In vitro-fertilized Bovine Follicular (가축의 개량 및 번식효율 증진에 관한 연구 Ⅵ. 소에 있어서 체외수정 난포란의 발생능 향상에 관한 연구)

  • 정영채;김창근;류범용;윤종택;김훈태;이규승
    • Korean Journal of Animal Reproduction
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    • v.14 no.1
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    • pp.73-83
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    • 1990
  • These studies were carried out to find the proper conditions for in vitro maturation and fertilization of bovine follicular oocytes and culture methods capable of further developing early embryos. For these objectives, the cleavage rate of oocytes matured and fertilized in vitro was investigated under medium supplemented with hormones and estrous cow serum and season of oocytes collection as well as different cumulus cell stage before insemination. Finally, 2~8 cell embryos were cultured in in vitro and in vitro culture system to investigate developmental capacity into morula. 1. Cleavage rate of oocytes matured in vitro was 27%(20/73) for A(LH+FSH+estradil-17$\beta$+10% FBS), 38%(27/71) for B(LH+10% ECS) and 27%(15/56) for C(10% ECS), respectively. Supplement B showed more higher rate and 4~8 cell embryos were also obtained much more in this group(67%, 18/27). In vitro maturation rate of follicular oocytes cultured in TCM 199 supplemented withLH and 10% ECS was 88%(75/85). 2. Cleavage rate(15%, 10/65) of oocytes collected in summer was significantly lower than in fall(47%, 42/89). 3. Cleavage rate(15%, 10/65) of oocytes collected in summer was significantly lower than in fall(47%, 42/89). 3. Cleavage rate(15%, 10/65) of oocytes with partially removed cumulus cell before insemination was more higher than that(44%, 27/62) of oocytes with intact cumulus cell. 4. The frequency of development from early cleaved embryos into morula was 6%(4/65), 12%(4/33) for co-culture of cumulus cell monolayer and bovine oviduct epithelial cell monolayer, respectively and 25%(6/25) in ligated rabbit oviduct.

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Extracellular 5-Aminolevulinic Acid Production by Escherichia coli Containing the Rhodopseudomonas palustris KUGB306 hemA Gene

  • Choi, Han-Pil;Lee, Young-Mi;Yun, Cheol-Won;Sung, Ha-Chin
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1136-1140
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    • 2008
  • The Rhodopseudomonas palustris KUGB306 hemA gene codes for 5-aminolevulinic acid (ALA) synthase. This enzyme catalyzes the condensation of glycine and succinyl-CoA to yield ALA in the presence of the cofactor pyridoxal 5'-phosphate. The R. palustris KUGB306 hemA gene in the pGEX-KG vector system was transformed into Escherichia coli BL21. The effects of physiological factors on the extracellular production of ALA by the recombinant E. coli were studied. Terrific Broth (TB) medium resulted in significantly higher cell growth and ALA production than did Luria-Bertani (LB) medium. ALA production was significantly enhanced by the addition of succinate together with glycine in the medium. Maximal ALA production (2.5 g/l) was observed upon the addition of D-glucose as an ALA dehydratase inhibitor in the late-log culture phase. Based on the results obtained from the shake-flask cultures, fermentation was carried out using the recombinant E. coli in TB medium, with the initial addition of 90 mM glycine and 120 mM succinate, and the addition of 45 mM D-glucose in the late-log phase. The extracellular production of ALA was also influenced by the pH of the culture broth. We maintained a pH of 6.5 in the fermenter throughout the culture process, achieving the maximal levels of extracellular ALA production (5.15 g/l, 39.3 mM).

Effects of Glucose on the Cleavage and Further Development of Early Bovine Embryos (Glucose가 소 초기배의 분할 및 발육에 미치는 영향)

  • 노상호;이병천;황우석
    • Journal of Embryo Transfer
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    • v.12 no.2
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    • pp.161-169
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    • 1997
  • This study was conducted to compare the insemination time of bovine oocytes and determine the effects of glucose(1.5 mM) on the development of bovine embryos at early cleavage stage. Oocytes were matured for 24 h, followed by exposure to sperm and cultured in modified Tyrode's media drops or with bovine oviduct epithelial cell monolayer prepared in TCM199(BOECM). Insemination time and culture system were varied in each experiment. In experiment 1, to investigate the developmental capacity of bovine embryos after different time of exposure to sperm, bovine ova and sperm were co-incubated for 18, 30 or 54 h, respectively. The development to blastocysts of 30 and 54 h insemination groups were significantly higher(P<0.05) than 18 h group, and in case of blastocysts of cleaved embryos, 30 h group were significantly higher(P<0.05) than other groups. In experiment 2, we investigated the effect of glucose on early bovine embryos. After 18 h insemination, in vitro fertilized oocytes were separated following 3 groups ; G+0, C+24 and C+48. Oocytes of G+0 group were cultured in glucose added Tyrode's medium after fertilization, oocytes in C+24 and C+48 groups were cultured in glucose free Tyrode's medium after fertilization. After 24 h culture, G+24 group was moved to glucose added medium. All oocytes of 3 groups were moved to BOECM after 48 h culture. The rates of cleavage and development to blastocysts in G+0 group were significantly lower than other groups. In experiment 3, we determined the effects of glucose exposure from 8 to 20 h after insemination on the cleavage and development of oocytes. The oocytes in glucose added group had high capacity of cleavage and further development. This study shows that in bovine oocytes, the optimal exposure to sperm is 30 h and glucose exposure to bovine one-cell embryos is detrimental to their first cleavage and further development in vitro but there has no evidence of detrimental effect of glucose(1.5 mM) exposure to bovine embryos over the two-cell stage in vitro.

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Genomic and Transgenic Approaches to Modified Plants: Disease Resistance in the Brassica as a Model System.

  • Ekuere, Usukuma;Good, Allen G.;Mayerhofer, Reinhold
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.4
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    • pp.317-323
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    • 2000
  • Molecular genetic techniques can now be applied to the development of advanced plant genotypes, either through genetic transformation or genomic approaches which allow researchers to transfer specific traits using molecular markers. In this paper, we discuss the use of these techniques towards understanding the genetics of blackleg resistance in Brassica. In a comparative mapping study between Arabidopsis thaliana and Brassica napus, 6 R-ESTs, 7 B. napus RFLP markers and a B. napus EST were located in a collinear region of N7 (B. napus) and chromosome 1 (A. thaliana). One of the A. thaliana R-ESTs and 4 of the B. napus RFLPs co-segregated and mapped to the LmRl locus for blackleg resistance. Introgression of blackleg resistance from wild relatives is also investigated with the possibility of accelerating the introgression process via marker assisted selection.

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Project Management based Analysis for the Enterprise 6 Sigma Success Factors (프로젝트 경영관점에서의 전사 6 시그마 성공요인 분석)

  • Mhun, Young-Soo;Bae, Suk-Joo
    • Journal of Applied Reliability
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    • v.11 no.1
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    • pp.59-81
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    • 2011
  • Six Sigma has been disseminated into a variety of areas. It is believed that a stepwise understanding of Six Sigma's DMAIC is required for successes of unit projects. Meanwhile, infra-structure factors, such as leadership, organization, personnel management, educational system, etc., are also important to safely implant the Six Sigma into the whole enterprise and to thoroughly create the culture of innovation and enterprise. Careful analysis of those elements makes it possible to draw common connection between Project Management and the Six Sigma. This study drives the success elements for the successful adoption of the Six Sigma in the Korea Western Power Co., Ltd., of which management was evaluated as one of the tops based on the results of 2007 Management Innovation Assessment and arranges the similarity of like this factor and Enterprise Project Management. The results of this study are expected to provide a useful guideline for the adoption of the Six Sigma in whole company level.

A Study on the Effect of the Mega trend on the Modern Fashion Design (현대 패션디자인에 나타난 메가 트렌드의 영향에 관한 연구)

  • Jun, Hye-Jung;Ha, Ji-Soo
    • Journal of the Korean Society of Costume
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    • v.57 no.4 s.113
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    • pp.110-125
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    • 2007
  • The purpose of this study is to analyze efforts of the mesa trends on the modern fashion design. For the study documentary research and qualitative content analysis have been carried out. Articles in magazines, seminar reports, and news related to trends were collected and analyzed to develop mega trends nowadays. As results, $2002{\sim}2006$ mega trends were extracted to Co-divergence, Emotional tech, Bi-dentity, Private+One and $2002{\sim}2006$ design trends extracted to Camouflage, Transformation, Modular system, Blur, Ease. The influences of mega trends and design trends on the modern fashion design have been proved. Those were expressed as eclectic style, deconstructionist style, reductive style and functionalist style in fashion design. The present era could be defined as cultural pluralism blended with various cultures and styles. Fashion as the essential display of contemporary culture and society has been successfully confirmed in this study.

유전독성물질의 평가방법과 그 기작에 관한 연구

  • 이정섭;박종근;박종광;박상대
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.170-170
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    • 1993
  • 유전독성물절외 검출과 평가에 용이하게 사용할 수 있는 모델 시스템의 개발 및 DNA 회복기작을 규명할 목적으로 수종의 돌연변이, 발암원을 이용하여 배양 포유동물세포 및 어류세포에서 세포생존률, DNA 합성 및 복제억제의 양상 등을 비교 분석하였다. MMS및 MNNG 와 같은 알칼라제는 CHO 세포에서 유의한 DNA 합성저해, DNA 복재억제, DNA 단사절단 및 비주기성 DNA 합성률의 증가를 유발하였다. Benzo(a)pyrene과 3-methylcholanthrene좌 같은 DNA 상해 전구물질의 경우 유전독성 여부의 판정에는 반드시 S-9/15과 같은 대사활성계 또는 mouse embryonic fibroblast와 같은 대사 활성능이 있는 세포와의 co-culture system들이 필요함을 확인하였으며, 이들에 의한 DNA 상해와 복재억제 유도의 작용양상은 자외선의 작용양상과 유사하였다. 배양 어류세포에서 자외선에 의한 세포생존율의 측정, 광재활성능의 분석 및 자외선에 의해 유발된 피리미딘 이량체 절제능 검토 및 DNA 합성 저해능의 결과를 분석함으로써 유전독성 평가를 위한 모델 시스템 구축의 기초결과를 얻었다.

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Effects of Ethylene and $Ca^{2+}$ on Activity of Phenylalanine Ammonia-Lyase in Glucan-Treated Daucus carota

  • Myoung-Won Kim
    • Journal of Plant Biology
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    • v.37 no.3
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    • pp.263-269
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    • 1994
  • Involvement of ethylene and Ca2+ on the induction of phenylalanine ammonia-lyase (PAL) was investigated in Daucus carota L. suspension culture system. Ethylene production started to increase about 3 h after glucan treatment. And the maximal induction of ethylene was preceded by PAL induction by 30 min. After the treatment of ethrel, PAL activity was increased. When cells were treated with glucan and Co2+, PAL activity was simultaneously reduced. Ethylene production was reduced dramatically in calcium-free medium, even though glucan was treated. PAL activity and ethylene producton was inhibited conspicuously when ethylene glycolbis($\beta$-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) was treated with glucan. Verapamil and trifluoperazine also inhibited PAL activity. When cells were treated with calcium ionophore A23187, PAL activity was increased in nontreated medium. We report here PAl activity is increased in related to ethylene production and involvement of Ca2+ in glucan-treated carrot suspension cells.

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Design of Content Scrambling System for DVB-IPTV Services using MPEG2 TS (MPEG2 TS 기반 DVB-IPTV 서비스를 위한 콘텐츠 스크램블링 시스템 설계)

  • Lee, Hyung;Shin, Jin-Seob;Im, Han-Jae
    • Proceedings of the Korean Society of Computer Information Conference
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    • 2017.07a
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    • pp.105-106
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    • 2017
  • 최근 고품질 방송콘텐츠 보호에 대한 요구가 증가하면서 DVB, MPEG, ATSC 등 국제표준단체에서는 보안 성능이 우수한 디지털방송 보호규격으로써 DVB-CSA ver.3에 대한 논의가 본격화되었다. 본 논문에서는 디지털방송에서 보안성능이 우수한 것으로 알려진 방송콘텐츠 암호화 규격인 DVB-CSA ver. 3 표준규격을 기반으로한 방송콘텐츠 스크램블링 시스템 설계에 대해 소개한다.

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