• Title/Summary/Keyword: cell line development

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

  • 제훈성;최차용
    • KSBB Journal
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    • v.7 no.2
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    • pp.96-101
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    • 1992
  • A low serum medium(LSM) suitable for the growth of a self-constructed hybridoma cell line, KA112, was established by selecting ingredients to replace serum. Insulin and sodium pyruvate was important for the growth of cell line KA112. Various basal media were tested and DMEM gave the most favorable result. Low serum medium (LSM) developed in this work showed cell line stability in the culture for more than 6 months and exhibited cell growth equivalent to that carried out in medium supplemented with 7% FBS. LSM was found to be applicable to the suspension culture of KA112. The reduction of serum level down to 1%(V/V) FBS in LSM resulted in a substantial saving in the cost of media preparation for large scale culture.

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

  • Park, Chungmu;Yoon, Hyunseo
    • Journal of The Korean Society of Integrative Medicine
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    • v.7 no.4
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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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    • v.13 no.10
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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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    • v.7 no.6
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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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    • v.1 no.1
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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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    • v.28 no.1
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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.

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 (폐암세포에 p16 (MTS1) 유전자 주입후 암생성능의 변화 및 세포주기관련 단백질의 변동에 관한 연구)

  • Kim, Young-Whan;Kim, Jae-Yeol;Yoo, Chul-Gyu;Han, Sung-Koo;Shim, Young-Soo;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.796-805
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    • 1997
  • Background : It is clear that deregulation of cell cycle progression is a hallmark of neoplastic transformation and genes involved in the $G_1$/S transition of the cell cycle are especially frequent targets for mutations in human cancers, including lung cancer. p16 gene product, one of the G1 cell-cycle related proteins, that is recently identified plays an important role in the negative regulation of the the kinase activity of the cyclin dependent kinase (cdk) enzymes. Therefore p16 gene is known to be an important tumor suppressor gene and is also called MTS1 (multiple tumor suppressor 1). No more oncogenes have been reported to be frequently related to multiple different malignancies than the alterations of p16 gene. Especially when it comes to non-small cell lung cancer, there was no expression of p16 in more than 70% of cell lines examined. And also it is speculated that p16 gene could exert a key role in the development of non-small cell lung cancer. This study was designed to evaluate whether p16 gene could be used as a candidate for gene therapy of non-small cell lung cancer. Methods : After the extraction of total RNA from normal fibroblast cell line and subsequent reverse transcriptase reaction and polymerase chain reaction, the amplified p16 cDNA was subcloned into eukaryotic expression plasmid vector, pRC-CMV. The constructed pRC-CMV-p16 was transfected into the NCI-H441 NSCLC cell line using lipofectin. The changes of G1 cell-cycle related proteins were investigated with Western blot analysis and immunoprecipitation after extraction of proteins from cell lysates and tumor suppressive effect was observed by clonogenic assay. Results : (1) p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 showed the formation of p16 : cdk 4 complex and decreased phosphorylated Rb protein, while control cell line did not. (2) Clonogenic assay demonstrated that the number of colony formation was markedly decreased in p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 than the control cell line. Conclusion : It is confirmed that the expression of p16 protein in p16 absent NSCLC cell line with the gene transfection leads to p16 : cdk4 complex formation, subsequent decrease of phosphorylated pRb protein and ultimately tumor suppressive effects. And also it provides the foundation for the application of p16 gene as a important candidate for the gene therapy of NSCLC.

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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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    • v.28 no.6
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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
    • Journal of the Korean Magnetic Resonance Society
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    • v.18 no.1
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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.