• Title/Summary/Keyword: cell line development

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Hybridoma 배양을 위한 저혈청 배지의 개발 제2부: 혈청 대체 물질 선정을 통한 저혈청 배지 제조 (Develpment of Low-serum Medium(LSM) for Mouse-mouse Hybridoma Part II. Development of Low Serum Medium by Screening for Serum Replacement)

  • 제훈성;최차용
    • KSBB Journal
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    • 제7권2호
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    • pp.96-101
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    • 1992
  • 저혈청 배지의 개발을 위해 혈청 대체 가능성이 있는 물질을 선정하여 각각의 필요성을 검색하였다. 검색 결과 결정된 물질들로 저혈청 배지를 제조하였으며, 기저 배지의 종류와 혼합에 따른 영향을 고찰 하였다. 개발된 배지는 1% FBS 첨가만으로 6개월 이상의 계대배양에서 만족할만한 안정성을 보였으며, 부유배양에도 적합한 것으로 판단되었다. 개발된 배지에서 혈청은 필수적이었으나 균주의 장기적응후에 무혈청 배지의 개발이 보다 용이할 것으로 기대되었다.

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사람 치은섬유모세포에서 잎꼬시래기 에탄올 추출물의 항염증 및 항산화 효과 (Anti-Inflammatory and Antioxidative Effects of Gracilaria textorii Ethanol Extract in LPS-PG-Stimulated Human Gingival Fibroblast-1 Cells)

  • 박충무;윤현서
    • 대한통합의학회지
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    • 제7권4호
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    • pp.61-69
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    • 2019
  • Purpose : Human gingival fibroblast cell is one of the the main cell types in periodontal tissue, which they can show anti-inflammatory activity through the production of numerous lines of inflammatory mediators such as inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and interleukins. Porphyromonas gingivalis, one of the oral pathogens, has reported to play a critical role in the development of periodontal diseases. This study aimed to investigate anti-inflammatory and antioxidative activities of Gracilaria textorii ethanol extract (GTEE) in P. gingivalis derived lipopolysaccharide (LPS-PG) stimulated human gingival fibroblast (HGF)-1 cell line. Methods : In order to analyze anti-inflammatory and antioxidative activities of GTEE in HGF-1 cell line, NOS enzyme activity, expression levels of iNOS, COX-2, NAD(P)H quinone dehydrogenase (NQO)1 and their transcription factors were estimated by Griess reaction and western hybridization. Results : LPS-PG induced overexpression of iNOS and COX-2, which was significantly attenuated by GTEE treatment in a dose-dependent manner without any cytotoxicity. In addition, intracellular NOS activity was in accordance with the result of iNOS expression. Due to important role in the regulation of inflammatory responses, phosphorylated status of p65 and c-jun, each subunit of nuclear factor (NF)-κB and activator protein (AP)-1, was also dose-dependently ameliorated by GTEE treatment. One of phase II enzymes, NQO1, and its transcription factor, nuclear factor erythroid 2-related factor 2 (Nrf2), were analyzed since elevated phase II enzyme expression inhibited inflammatory response, which was significantly elevated by GTEE treatment in HGF-1 cell line. Conclusion : In conclusion, GTEE mitigated LPS-PG-stimulated inflammatory responses by attenuating NF-κB and AP-1 activation as well as accelerating NQO1 and Nrf2 expression in HGF-1 cell line. These results indicate that GTEE might be utilized a promising strategy for potential anti-inflammatory agent in periodontal diseases.

Synaptic Vesicle Protein 2 (SV2) Isoforms

  • Bandala, Cindy;Miliar-Garcia, A.;Mejia-Barradas, C.M.;Anaya-Ruiz, M.;Luna-Arias, J.P.;Bazan-Mendez, C.I.;Gomez-Lopez, M.;Juarez-Mendez, S.;Lara-Padilla, E.
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권10호
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    • pp.5063-5067
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    • 2012
  • New molecular markers of cancer had emerged with novel applications in cancer prevention and therapeutics, including for breast cancer of unknown causes, which has a high impact on the health of women worldwide. The purpose of this research was to detemine protein and mRNA expression of synaptic vesicle 2 (SV2) isoforms A, B and C in breast cancer cell lines. Cultured cell lines MDA-MB-231, SKBR3, T47D were lysed and their protein and mRNA expression analyzed by real-time PCR and western blot technique, respectively. SV2A, B proteins were identified in non-tumor (MCF-10A) and tumor cell lines (MDA-MB-231 and T47D) while SV2C only was found in the T47D cell line. Furthermore, the genomic expression was consistent with protein expression for a such cell line, but in MDA-MB-231 there was no SV2B genomic expression, and the SV2C mRNA and protein were not found in the non tumoral cell line. These findings suggest a possible cellular transdifferentiation to neural character in breast cancer, of possible relevance to cancer development, and point to possible use of SV2 as molecular marker and a vehicle for cancer treatment with botulinum toxin.

DEVELOPMENT OF DIGITAL ASSEMBLY CELL FOR LASER WELDING OF SIDE PANELS

  • Park, H.S.;Lee, H.B.
    • International Journal of Automotive Technology
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    • 제7권6호
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    • pp.721-728
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    • 2006
  • This paper is concerned with development of digital assembly cell. Automotive industries, try to find out new welding technologies to replace the existing spot welding which is not appropriate anymore. Because of many advantages like good accessibility, welding quality and fast welding speed, laser welding seems the most appropriate solution for automobile manufactures. To apply this technology to welding of car body, experiments must be conducted according to material, geometry and layer in welding area. Based on the data of these experiments, the laser welding process and the quality of stitches are identified. And then, the comparison between the requirements of welding and the potential of equipments allows selecting the best equipments. By using the digital manufacturing, the configurations of laser welding cell are carried out. After all, the optimal laser welding cell is chosen by the evaluation of alternative cells with technical and organizational criteria.

A SDR/DDR 4Gb DRAM with $0.11\mu\textrm{m}$ DRAM Technology

  • Kim, Ki-Nam
    • JSTS:Journal of Semiconductor Technology and Science
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    • 제1권1호
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    • pp.20-30
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    • 2001
  • A 1.8V $650{\;}\textrm{mm}^2$ 4Gb DRAM having $0.10{\;}\mu\textrm{m}^2$ cell size has been successfully developed using 0.11 $\mu\textrm{m}$DRAM technology. Considering manufactur-ability, we have focused on developing patterning technology using KrF lithography that makes $0.11{\;}\mu\textrm{m}$ DRAM technology possible. Furthermore, we developed novel DRAM technologies, which will have strong influence on the future DRAM integration. These are novel oxide gap-filling, W-bit line with stud contact for borderless metal contact, line-type storage node self-aligned contact (SAC), mechanically stable metal-insulator-silicon (MIS) capacitor and CVD Al process for metal inter-connections. In addition, 80 nm array transistor and sub-80 nm memory cell contact are also developed for high functional yield as well as chip performance. Many issues which large sized chip often faces are solved by novel design approaches such as skew minimizing technique, gain control pre-sensing scheme and bit line calibration scheme.

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A Mutation of cdc-25.1 Causes Defects in Germ Cells But Not in Somatic Tissues in C. elegans

  • Kim, Jiyoung;Lee, Ah-Reum;Kawasaki, Ichiro;Strome, Susan;Shim, Yhong-Hee
    • Molecules and Cells
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    • 제28권1호
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    • pp.43-48
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    • 2009
  • By screening C. elegans mutants for severe defects in germline proliferation, we isolated a new loss-of-function allele of cdc-25.1, bn115. bn115 and another previously identified loss-of-function allele nr2036 do not exhibit noticeable cell division defects in the somatic tissues but have reduced numbers of germ cells and are sterile, indicating that cdc-25.1 functions predominantly in the germ line during postembryonic development, and that cdc-25.1 activity is probably not required in somatic lineages during larval development. We analyzed cell division of germ cells and somatic tissues in bn115 homozygotes with germline-specific anti-PGL-1 immunofluorescence and GFP transgenes that express in intestinal cells, in distal tip cells, and in gonadal sheath cells, respectively. We also analyzed the expression pattern of cdc-25.1 with conventional and quantitative RT-PCR. In the presence of three other family members of cdc-25.1 in C. elegans, defects are observed only in the germ line but not in the somatic tissues in cdc-25.1 single mutants, and cdc-25.1 is expressed predominantly, if not exclusively, in the germ line during postembryonic stages. Our findings indicate that the function of cdc-25.1 is unique in the germ line but likely redundant with other members in the soma.

폐암세포에 p16 (MTS1) 유전자 주입후 암생성능의 변화 및 세포주기관련 단백질의 변동에 관한 연구 (The Change of Cell-cycle Related Proteins and Tumor Suppressive Effect in Non-small Cell Lung Cancer Cell Line after Transfection of p16(MTS1) Gene)

  • 김영환;김재열;유철규;한성구;심영수;이계영
    • Tuberculosis and Respiratory Diseases
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    • 제44권4호
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    • pp.796-805
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    • 1997
  • 연구배경 : 세포주기의 활성화, 그 중에서도 특히 $G_1$/S 이행에 관여하는 세포주기관련 단백질들은 암발생에 있어서 매우 중요한 역할을 하는 것으로 알려져 있다. $G_1$ 세포주기 관련 단백질 중의 하나인 cdk4 (cyclin dependent kinase 4)의 억제제로 알려져 있는 p16 유전자는 최근에 밝혀진 종양억제유전자중의 하나로서 MTS1 (multiple tumor suppressor 1)이라고도 불린다. p16 유전자는 지금까지 알려진 어느 종양관련 유전자보다도 유전자변이의 빈도가 높은 암억제유전자인데, 특히 비소세포폐암인 경우는 70% 이상의 세포주에서 p16 단백질의 발현이 없는 것으로 밝혀져 있어 p16 유전자는 비소세포폐암 발생에 매우 중요한 역할을 할 것이라고 알려져 있다. 본 연구에서는 비소세포폐암에서 p16을 이용한 유전자치료의 타당성을 입증하기 위하여 다음과 같은 연구를 시행하였다. 방 법 : p16이 결여된 비소세포폐암 세포주 (NCI-H441)에, 정상섬유아세포에서 총 RNA를 추출하여 역전사효소 및 DNA 중합효소반응으로 증폭된 p16 cDNA를 유핵세포 발현 vector인 pRC-CMV plasmid에 subcloning하여 구축된 pRC-CMV-p16 plasmid vector를 lipofectin을 이용하여 유전자 이입한 후, 단백질을 추출하여 Western blot 분석과 면역침전법으로 $G_1$ 세포주기관련 단백질의 변동을 관찰하고, colony 형성능을 비교함으로써 암억제효과를 확인하였다. 결 과 : p16이 유전자주입된 NCI-H441 세포주에서 p16과 cdk4가 복합체를 형성하고 있고 인산화 Rb가 대조 세포주에 비해 감소되어 있음을 확인할 수 있어, p16이 cdk4와 결합함으로써 cdk4에 의한 Rb의 인산화를 방해하고 이에 따른 $G_1$ 세포주기 정체에 의해 종양억제효과가 나타난다는 설명을 뒷받침할 수 있었다. Clonogenic assay 결과는 p16 유전자주입된 NCI-H441 세포주의 colony 형성능이 대조 세포주에 비하여 현격히 감소함을 관찰하였다. 결 론 : 이상의 결과로 p16(MTS1) 유전자를 p16 단백질을 발현하지 못하는 비소세포폐암 세포주에 주입할 경우, 주입한 유전자에서 생성되는 p16 단백질이 cdk와 결합하여 Rb 단백질의 인산화를 저하시켜 궁극적으로 암억제 효과를 일으킬 수 있음이 확인되었고, 이는 향후 비소세포폐암의 유전자치료에 있어서 p16 유전자의 이용 가능성을 확인한 기초자료가 된다고 생각된다.

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Twist2 Regulates CD7 Expression and Galectin-1-Induced Apoptosis in Mature T-Cells

  • Koh, Han Seok;Lee, Changjin;Lee, Kwang Soo;Park, Eun Jung;Seong, Rho H.;Hong, Seokmann;Jeon, Sung Ho
    • Molecules and Cells
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    • 제28권6호
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    • pp.553-558
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    • 2009
  • In the periphery, a galectin-1 receptor, CD7, plays crucial roles in galectin-1-mediated apoptosis of activated T-cells as well as progression of T-lymphoma. Previously, we demonstrated that $NF-{\kappa}B$ downregulated CD7 gene expression through the p38 MAPK pathway in developing immature thymocytes. However, its regulatory pathway is not well understood in functional mature T-cells. Here, we show that CD7 expression was downregulated by Twist2 in Jurkat cells, a human acute T-cell lymphoma cell line, and in EL4 cells, a mature murine T-cell lymphoma cell line. Furthermore, ectopic expression of Twist2 in Jurkat cells reduced galectin-1-induced apoptosis. While full-length Twist2 decreased CD7 promoter activity, a C-terminal deletion form of Twist2 reversed its inhibition, suggesting an important role of the C-terminus in CD7 regulation. In addition, CD7 expression was enhanced by histone deacetylase inhibitors such as trichostatin A and sodium butyrate, which indicates that Twist2 might be one of candidate factors involved in histone deacetylation. Based on these results, we conclude that upregulation of Twist2 increases the resistance to galectin-1-mediated-apoptosis, which may have significant implications for the progression of some T-cells into tumors such as Sezary cells.

Metabolic perturbation of an Hsp90 C-domain inhibitor in a lung cancer cell line, A549 studied by NMR-based chemometric analysis

  • Hur, Su-Jung;Lee, Hye-Won;Shin, Ai-Hyang;Park, Sung Jean
    • 한국자기공명학회논문지
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    • 제18권1호
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    • pp.10-14
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    • 2014
  • Hsp90 is a good drug target molecule that is involved in regulating various signaling pathway in normal cell and the role of Hsp90 is highly emphasized especially in cancer cells. Thus, much efforts for discovery and development of Hsp90 inhibitor have been continued and a few Hsp90 inhibitors targeting the N-terminal ATP binding site are being tested in the clinical trials. There are no metabolic signature molecules that can be used to evaluate the effect of Hsp90 inhibition. We previously found a potential C-domain binder named PPC1 that is a synthetic small molecule. Here we report the metabolomics study to find signature metabolites upon treatment of PPC1 compound in lung cancer cell line, A549 and discuss the potentiality of metabolomic approach for evaluation of hit compounds.