• Title/Summary/Keyword: Cell division

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Screening of High-Productivity Cell Lines and Investigation of Their Physiology in Chinese Hamster Ovary (CHO) Cell Cultures for Transforming Growth $Factor-{\beta}1$ Production

  • Chun, Gin-Taek;Lee, Joo-Buom;Nam, Sang-Uk;Lee, Se-Won;Jeong, Yeon-Ho;Choi, Eui-Yul;Kim, Ik-Hwan;Jeong, Yong-Seob;Kim, Pyeong-Hyeun
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.121-129
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    • 2002
  • Using recombinant Chinese hamster ovary (CHO) cells, strategies for developing high producers for the recombinant human Transforming Growth $Factor-{\beta}1$ ($TGF-{\beta}1$) protein are proposed and their physiological characteristics in cell cultures were investigated. $TGF-{\beta}1$ is a pleiotrophic polypeptide involved in various biological activities, including cell growth, differentiation, and deposition of extracellular matrix proteins. The CHO cells included human $TGF-{\beta}1$ cDNA in conjunction with a dihydrofolate reductase (DHFR) gene, which was cotransfected into the cells to amplify the transfected $TGF-{\beta}1$ cDNA. As a first-round screening of the transfected cells, a relatively high $TGF-{\beta}1$-producing cell line was selected, and then, it acquired a resistance to increasing concentrations of methotrexate (MTX) up to $60{\mu}M$,resulting in a significant improvement in its $TGF-{\beta}1$ biosynthetic ability. After applying a monoclonal selection strategy to the MTX-resistant cells, more productive cells were screened, including the APP-3, App-5, and App-8 cell lines. These high producers were compared with two other cell lines (AP-l cell line without amplification of transfected $TGF-{\beta}1$ cDNA and nontransfectant of $TGF-{\beta}1$ cDNA) in terms of cell growth, $TGF-{\beta}1$ productivity, sugar uptake, and byproduct formation, in the presence or absence of MTX in the culture medium. Consequently, both monoclonal selection as well as an investigation of the physiological characteristics were found to be needed for the efficient screening of higher $TGF-{\beta}1$ producers, even after the transfection and amplification of the transfected gene.

Distributed Relay Power Control Scheme for Multi-cell OFDM-TDD Based Mobile Relay System (OFDM-TDD 기반 이동 릴레이 시스템을 위한 다중 셀 분산형 릴레이 전력 제어 방법)

  • Cho, Young-Min;Park, Jeong-Hun;Hwang, Seung-Gye;Kim, Dong-Ku
    • Journal of Advanced Navigation Technology
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    • v.15 no.4
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    • pp.562-570
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    • 2011
  • In this paper, a distributed mobile relay power control (DMRPC) scheme for maximizing individual cell throughput is proposed for mobile relay aided multi-cell orthogonal frequency division multiplexing (OFDM)-time division duplex (TDD) system. In the system with DMRPC, the power levels of relay's are controlled by individual cell without cell cooperation and signalling overhead. It is demonstrated by numerical simulation that DMRPC provides the better cell throughput performance than either the full power relay aided system or conventional system without relay does. Moreover, it is also shown that relay aided systems with DMRPC, and the conventional system have almost identical cell edge throughput, while full power relay aided systems show worse performance in cell edge throughput.

Monitoring the Differentiation and Migration Patterns of Neural Cells Derived from Human Embryonic Stem Cells Using a Microfluidic Culture System

  • Lee, Nayeon;Park, Jae Woo;Kim, Hyung Joon;Yeon, Ju Hun;Kwon, Jihye;Ko, Jung Jae;Oh, Seung-Hun;Kim, Hyun Sook;Kim, Aeri;Han, Baek Soo;Lee, Sang Chul;Jeon, Noo Li;Song, Jihwan
    • Molecules and Cells
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    • v.37 no.6
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    • pp.497-502
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    • 2014
  • Microfluidics can provide unique experimental tools to visualize the development of neural structures within a microscale device, which is followed by guidance of neurite growth in the axonal isolation compartment. We utilized microfluidics technology to monitor the differentiation and migration of neural cells derived from human embryonic stem cells (hESCs). We co-cultured hESCs with PA6 stromal cells, and isolated neural rosette-like structures, which subsequently formed neurospheres in suspension culture. Tuj1-positive neural cells, but not nestin-positive neural precursor cells (NPCs), were able to enter the microfluidics grooves (microchannels), suggesting that neural cell-migratory capacity was dependent upon neuronal differentiation stage. We also showed that bundles of axons formed and extended into the microchannels. Taken together, these results demonstrated that microfluidics technology can provide useful tools to study neurite outgrowth and axon guidance of neural cells, which are derived from human embryonic stem cells.

Stability Assessment of Lead Sulfide Colloidal Quantum Dot Based Schottky Solar Cell

  • Song, Jung-Hoon;Kim, Jun-Kwan;An, Hye-Jin;Choi, Hye-Kyoung;Jeong, So-Hee
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.08a
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    • pp.413-413
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    • 2012
  • Lead sulfide (PbS) Colloidal quantum dots (CQDs) are promising material for the photovoltaic device due to its various outstanding properties such as tunable band-gap, solution processability, and infrared absorption. More importantly, PbS CQDs have large exciton Bohr radius of 20 nm due to the uniquely large dielectric constants that result in the strong quantum confinement. To exploit desirable properties in photovoltaic device, it is essential to fabricate a device exhibiting stable performance. Unfortunately, the performance of PbS NQDs based Schottky solar cell is considerably degraded according to the exposure in the air. The air-exposed degradation originates on the oxidation of interface between PbS NQDS layer and metal electrode. Therefore, it is necessary to enhance the stability of Schottky junction device by inserting a passivation layer. We investigate the effect of insertion of passivation layer on the performance of Schottky junction solar cells using PbS NQDs with band-gap of 1.3 eV. Schottky solar cell is the simple photovoltaic device with junction between semiconducting layer and metal electrode which a significant built-in-potential is established due to the workfunction difference between two materials. Although the device without passivation layer significantly degraded in several hours, considerable enhancement of stability can be obtained by inserting the very thin LiF layer (<1 nm) as a passivation layer. In this study, LiF layer is inserted between PbS NQDs layer and metal as an interface passivation layer. From the results, we can conclude that employment of very thin LiF layer is effective to enhance the stability of Schottky junction solar cells. We believe that this passivation layer is applicable not only to the PbS NQDs based solar cell, but also the various NQDs materials in order to enhance the stability of the device.

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Standard Field Generation Using a Micro-TEM Cell and Its Measurement Uncertainty Evaluation (Micro-TEM Cell을 사용한 표준 전자기장의 발생 및 측정불확도 평가)

  • Kang, Jin-Seob;Kim, Jeong-Hwan;Kang, Ung-Taek;Kang, No-Weon;Kang, Tae-Weon
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.20 no.1
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    • pp.91-99
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    • 2009
  • In this paper, a standard field generation method using a micro-TEM ceil is described and its measurement uncertainty is evaluated. The standard field generation system consists of an auto-leveling signal source, a micro-TEM cell operating up to 1.2 GHz, and a power measuring Instrument using a thermistor mount. Measurement results of a field strength key comparison (CCEM.RF-K20) for the field strength of 20 V/m at frequencies between 10 MHz and 1 GHz are presented for validating the standard field generation method.

Effect of Activation Method and Culture Medium on the Development of Porcine Nuclear Transfer Embryo using Fetal Fibroblast

  • Im, Gi-Sun;Yang, Byoung-Chul;Park, Jin-Ki;Kim, Hyun-Ju;Chang, Won-Kyung;R. S. Prather;B. N. Day
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.66-66
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    • 2001
  • Since the first birth of pig derived from embryonic cells by nuclear transfer, many researches to produce cloned pig have been carried out. Recently, two reports about the birth of somatic cell cloned pigs using in vivo oocytes and also Betthauser et al. (2000) reported the birth of somatic cell cloned pigs using in vitro oocytes. So here we investigated the effect of activation method and culture medium on in vitro development of porcine nuclear transfer embryo using fetal fibroblast. Oocytes derived from slaughter house obtained ovaries were matured for 42 to 44 h in TCM 199. Matured oocytes were denuded using 0.1% hyaluronidase and then Oocytes with the first polar body were used for enucleation by aspirating the first polar body and adjacent cytoplasm in TCM 199 supplemented with 7.5 $\mu\textrm{g}$ cytochalasin B. Petal fibroblast cells were prepared from 35 days old fetus. To be used as donor cells, fetal fibroblast cells were serum starved for 3 to 5 days and then isolated into single co:1 by trypsinization. Nuclear transfer embryos were fused using 2 times 1.25㎸ for 30$mutextrm{s}$. Fused NT embryos were activated with calcium ionophore (CI) and 6-dimethyl-aminopurine (6-DMAP). Activated oocytes were cultured in NCSU 23 or BECM 3 for 6 days. There was no significant difference between chemical activation and no chemical activation for blastocyst development rate(11.6 vs. 14.8%). However, cell number was significantly higher when NT embryos were activated with CI and 6-DMAP (31.2 vs. 22.6). When NT embryos were cultured in NCSU 23 or BECM 3, blastocyst development rate was 16.4 and 13.2%, respectively, and cell number was 31.5 and 24.1, respectively. These results suggest that chemical activation after fusion and culture in NCSU 23 could increase cell number of porcine NT embryos.

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Effects of Cobalt Protective Coating Prepared by DC Electroplating on Ferritic Stainless Steel for SOFC Interconnect (직류 전기도금을 이용한 고체산화물 연료전지 금속연결재용 페라이트계 스테인리스 스틸의 코발트 보호막 코팅 효과)

  • Hong, Jong-Eun;Lim, Tak-Hyung;Song, Rak-Hyun;Lee, Seung-Bok;Shin, Dong-Ryul;Yoo, Young-Sung;Lee, Dok-Yol
    • Transactions of the Korean hydrogen and new energy society
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    • v.20 no.2
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    • pp.116-124
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    • 2009
  • We investigated the influences of cobalt coating deposited by DC electroplating on the ferritic stainless steel, STS 430, as a protective layer on a metallic interconnect for SOFC applications. Cobalt coated STS 430 revealed a uniform and denser-packing oxide surface and a reduced growth rate of $Cr_2O_3$ scales after oxidation at $800^{\circ}C$in air. Cobalt coating layer was oxidized to $CoCo_2O_4$ and Co containing mixed oxide spinels such as $Co_2CrO_4$, $CoCr_2O_4$, and $CoCrFeO_4$. The area specific resistance value of Co coated sample was $0.020\;{\Omega}cm^2$ lower than that of uncoated at $800^{\circ}C$ in air during 500 h. After 1000 h oxidation, cobalt oxide coating layer suppressed chromium outward diffusion.

Soluble CD30: A Possible Serum Tumor Marker for Primary Effusion Lymphoma

  • Michai, Manthana;Goto, Hiroki;Hattori, Shinichiro;Vaeteewoottacharn, Kulthida;Wongkham, Chaisiri;Wongkham, Sopit;Okada, Seiji
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.4939-4941
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    • 2012
  • Background: The serum level of soluble CD30 (sCD30) is known to be increased with several lymphomas and to correlate with prognosis. Primary effusion lymphoma (PEL) is a highly aggressive malignant lymphoma with poor prognosis, but the existence and significance of sCD30 in PEL have not yet been investigated in detail. Objectives: Since the membrane type of CD30 is frequently expressed on the surface of PEL cells, we compared the expression of the membrane type of CD30 and the production of sCD30 among PEL cell lines as well as other lymphomas. Methods: The expression of surface CD30 in various lymphoma cell lines was analyzed with flow cytometry ans sCD30 was quantified by ELISA. Results: Both surface and sCD30 were detected on PEL cell lines as well as on Hodgkin's lymphoma and adult T-cell leukemia/lymphoma cell lines. Surface CD30 and sCD30 levels of each cell lines correlated with each other. Conclusion: The serum level of sCD30 appear to be a useful biological tumor marker for the diagnosis and management of CD30-positive PEL.

N-Acetyl-D-Glucosamine Kinase Interacts with Dynein-Lis1-NudE1 Complex and Regulates Cell Division

  • Sharif, Syeda Ridita;Islam, Md. Ariful;Moon, Il Soo
    • Molecules and Cells
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    • v.39 no.9
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    • pp.669-679
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    • 2016
  • N-acetyl-D-glucosamine kinase (GlcNAc kinase or NAGK) primarily catalyzes phosphoryl transfer to GlcNAc during amino sugar metabolism. Recently, it was shown NAGK interacts with dynein light chain roadblock type 1 (DYNLRB1) and upregulates axo-dendritic growth, which is an enzyme activity-independent, non-canonical structural role. The authors examined the distributions of NAGK and NAGK-dynein complexes during the cell cycle in HEK293T cells. NAGK was expressed throughout different stages of cell division and immunocytochemistry (ICC) showed NAGK was localized at nuclear envelope, spindle microtubules (MTs), and kinetochores (KTs). A proximity ligation assay (PLA) for NAGK and DYNLRB1 revealed NAGK-dynein complex on nuclear envelopes in prophase cells and on chromosomes in metaphase cells. NAGK-DYNLRB1 PLA followed by Lis1/NudE1 immunostaining showed NAGK-dynein complexes were colocalized with Lis1 and NudE1 signals, and PLA for NAGK-Lis1 showed similar signal patterns, suggesting a functional link between NAGK and dynein-Lis1 complex. Subsequently, NAGK-dynein complexes were found in KTs and on nuclear membranes where KTs were marked with CENP-B ICC and nuclear membrane with lamin ICC. Furthermore, knockdown of NAGK by small hairpin (sh) RNA was found to delay cell division. These results indicate that the NAGK-dynein interaction with the involvements of Lis1 and NudE1 plays an important role in prophase nuclear envelope breakdown (NEB) and metaphase MT-KT attachment during eukaryotic cell division.

Dual Inhibition of PI3K/Akt/mTOR Pathway and Role of Autophagy in Non-Small Cell Lung Cancer Cells

  • Jeong, Eun-Hui;Choi, Hyeong-Sim;Lee, Tae-Gul;Kim, Hye-Ryoun;Kim, Cheol-Hyeon
    • Tuberculosis and Respiratory Diseases
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    • v.72 no.4
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    • pp.343-351
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    • 2012
  • Background: The phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling axis has emerged as a novel target for cancer therapy. Agents that inhibit this pathway are currently under development for lung cancer treatment. In the present study, we have tested whether dual inhibition of PI3K/Akt/mTOR signaling can lead to enahnced antitumor effects. We have also examined the role of autophagy during this process. Methods: We analyzed the combination effect of the mTOR inhibitor, temsirolimus, and the Akt inhibitor, GSK690693, on the survival of NCI-H460 and A549 non-small cell lung cancer cells. Cell proliferation was determined by MTT assay and apoptosis induction was evaluated by flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Autophagy induction was also evaluated by acridine orange staining. Changes of apoptosis or autophagy-related proteins were evaluated by western blot analysis. Results: Combination treatment with temsirolimus and GSK690693 caused synergistically increased cell death in NCI-H460 and A549 cells. This was attributable to increased induction of apoptosis. Caspase 3 activation and poly(ADP-ribose) polymerase cleavage accompanied these findings. Autophagy also increased and inhibition of autophagy resulted in increased cell death, suggesting its cytoprotective role during this process. Conclusion: Taken together, our results suggest that the combination of temsirolimus and GSK690693 could be a novel strategy for lung cancer therapy. Inhibition of autophagy could also be a promising method of enhancing the combination effect of these drugs.