• Title/Summary/Keyword: cell aggregation

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Effects of Storage-protein 2 Derived from Silkworm Hemolymph on Reduction of Aggregation and Cell Death in CHO Cells (CHO 세포에서 누에 혈림프 유래 Storage-protein 2의 세포응집 및 세포사멸 억제 효과)

  • Lim, Jin-Hyuk;Cha, Hyun-Myoung;Kim, Z-Hun;Choi, Yong-Soo;Kim, Dong-Il
    • KSBB Journal
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    • v.31 no.1
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    • pp.66-72
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    • 2016
  • Chinese hamster ovary (CHO) cells have been widely used for production of various recombinant proteins such as cytokines and monoclonal antibodies. The cell aggregation and cell death in CHO cell culture directly affect cell viability, and productivity and quality of products. In this study, we investigated preventing effects of storage-protein 2 (SP2) derived from silkworm hemolymph on cell aggregation and cell death in CHO cell culture producing albuminerythropoietin (Alb-EPO). The viable cell density in the culture supplemented with 2 mg/mL SP2 was 1.71-fold higher than that in control culture. Increased titer of Alb-EPO was also found in the culture with SP2. Morphology of CHO cells in SP2 supplemented cultures did not differ from that of control. In addition, the cell aggregation rate of the SP2 cultures was reduced 20% compared to the control. Finally, we confirmed that the apoptosis was strongly suppressed by addition of SP2 in the cultures. These results clearly demonstrate that SP2 can be served as an effective supplement for enhancing titer of Alb-EPO via reducing cell aggregation and cell death.

Ultrasound Backscattering from Erythrocyte Aggregation of Human, Horse and Rat Blood under Rotational Flow in a Cylindrical Chamber

  • Nam, Kweon-Ho;Paeng, Dong-Guk;Choi, Min-Joo
    • The Journal of the Acoustical Society of Korea
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    • v.25 no.4E
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    • pp.159-165
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    • 2006
  • Human, horse and rat bloods in a cylindrical chamber where flow was controlled by a stirring magnet were used for studying red blood cell aggregation. Ultrasound backscattered powers from blood were obtained from the backscattered signals measured by a 5 MHz focused transducer in a pulse-echo setup. The experimental results showed the differences in red blood cell (RBC) aggregation tendency among the three mammalian species with an order of horse > human > rat. The ultrasound backscattered power decreased with stirring speed in human and horse blood, but no variations were observed in rat blood. Sudden flow stoppage led to the slow increase of the backscattered power for human and horse blood. There was no self-aggregation tendency in rat blood. The enveloped echo images showed the spatial and temporal variations of RBC aggregations in the cylindrical chamber. These observations from the different mammalian species may give a better understanding of the mechanism of RBC aggregation.

The Effect of Transverse Vibration on Red Blood Cell Aggregation and Blood Viscosity

  • Shin, Se-Hyun;Ku, Yun-Hee;Park, Myung-Su;Suh, Jang-Soo
    • International Journal of Vascular Biomedical Engineering
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    • v.1 no.2
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    • pp.4-12
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    • 2003
  • The present study investigated the effect of transverse vibration on the hemorheological characteristics of blood using a newly designed pressure-scanning capillary viscometer. As vibration was applied, aggregated blood cells (rouleaux) were disaggregated. The range of vibration frequency and amplitude are $0{\sim}100\;Hz$ and $0{\sim}0.8\;mm$, respectively for a capillary diameter 0.84 mm. As vibration increased, blood viscosity initially increased and tended to decrease. In order to delineate the unexpected results, the present study proposed two counteracting mechanisms of vibration related with red blood cell (RBC) aggregation affecting hemo-rheological properties. One is the reduction of RBC aggregation due to vibration causing an increase of blood viscosity. The other is forced cell migration due to the transverse vibration, which in turn forms a cell-free layer near the tube wall and causes a decrease of flow resistance.

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Production of a Platelet Aggregation Inhibitor, Salmosin, by High Cell Density Fermentation of Recombinant Escherichia coli

  • Seo, Myung-Ji;Choi, Hak-Jong;Chung, Kwang-Hoe;Pyun, Yu-Ryang
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1053-1056
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    • 2011
  • Optimal conditions for a high cell density fermentation were investigated in a recombinant Escherichia coli producing salmosin, a platelet aggregation inhibitor. The optimized carbon and nitrogen sources were glycerol 10 g/l, yeast extract 30 g/l, and bacto-tryptone 10 g/l, yielding the dry cell weight (DCW) of 10.61 g/l in a 500 ml flask culture. The late-stage induction with 1% L-arabinose in a 5 l jar fermentor showed the highest DCW of 65.70 g/l after 27 h of the fed-batch fermentation. Around 2,200 mg/l of the protein was expressed as an inclusion body that was then refolded to obtain the active salmosin of 96 mg/l. We also confirmed the inhibitory activity against platelet aggregation of the active salmosin from the high cell density fermentation.

Effect of Cell Aggregation on Camptothecin Production in Suspension Cultures of Camptotheca acuminata (Camptotheca acuminata 현탁배양에서 세포응집이 Camptothecin 생산에 미치는 영향)

  • Choi, Hoon;Byun, Sang-Yo
    • KSBB Journal
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    • v.15 no.6
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    • pp.609-614
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    • 2000
  • Suspension cultures of Camptotheca acuminata, which is known to produce the anticancer indole alkaloid camptothecin and its derivatives, were made to increase camptothecin production. The capability of camptothecin production in suspended cells is decreased by repeated subculturein. Aggregated cells produced more camptothecin than single cells. Optimal cell aggregation was achieved in hybrid medium supplemented with 4% sucrose. Aggregated cells in hybrid medium with 4% sucrose produced $18.04{\times}10^{-4} mg/L$ of camptothecin. The control of shaking speeds was effective at inducing cell aggregation and camptothecin production. A shaking speed of 100 rpm was found optimum to increase the cell aggregation with a camptothecin production of $19.4{\times}10^{-4} mg/L$.

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Measurement of cell aggregation characteristics by analysis of laser-backscattering in a microfluidic rheometry

  • Shin, Se-Hyun;Hou, J.X.;Suh, Jang-Soo
    • Korea-Australia Rheology Journal
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    • v.19 no.2
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    • pp.61-66
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    • 2007
  • The aggregation characteristics of red blood cells (RBCs) are known as important factors in the microvascular flow system, and increased RBC aggregation has been observed in various pathological diseases, such as thrombosis and myocardial infarction. This paper describes a simple microfluidic device for measuring the RBC aggregation by integrating a microfluidic slit rheometry and laser-backscattering technique. While a decreasing-pressure mechanism was applied to the microfluidic rheometry, a syllectogram (the light intensity versus time) showed an initial increase and a peak caused by the high shear stress-induced disaggregation, immediately followed by a decrease in the light intensity due to RBC aggregation. The critical shear stress (CST) corresponding to the peak intensity was examined as a new index of the RBC aggregation characteristics. The CST of RBCs increased with increasing aggregation-dominating protein (fibrinogen) in the blood plasma. The essential feature of this design was the combination of the rheometric-optic characterization of RBC aggregation with a microfluidic chip, which may potentially allow cell aggregation measurements to be easily carried out in a clinical setting.

In Vitro Aggregation and Culture of Mouse Embryos (생쥐 배의 시험관내 응집과 배양)

  • 이상진;정길생
    • Korean Journal of Animal Reproduction
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    • v.8 no.1
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    • pp.29-35
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    • 1984
  • These experiments were carried out to obtain basic information necessary for in vitro culture of aggregated mouse embryos. Inbred ICR mice were used to obtain embryos. The zona pellucida was removed by placing the embryos in Whittingham's medium containing 0.5% protease for about 5-10minutes at 37$^{\circ}C$. Total 263 pairs of 2-, 4- and 8-cell zona free mouse embryos were subjected to aggregation by physical pressure and cultured in Whittingham's medium under the gas phase of 5% CO2 in air at 37$^{\circ}C$ for 24 to 60 hours. The results obtained in these experiments were summarized as follows: 1. Time needed for fusion of 2-, 4- and 8-cell embryos were 0-3, 0-3 and 0-3 hours, respectively and average time needed for in vitro development of 2-, 4- and 8-cell embryos after aggregation to morula and blastocyst were 42, 30 and 13.5 hours, and 51, 39 and 27 hours, respectively. 2. Of total 263 pairs of naked embryos, 227 were firmly aggregated together and the rats of aggregation in 2-, 4- and 8-cell embryos were 71.8, 88.3 and 97.0%, respectively. 3. The rates of aggregated pairs which obtained from 2-, 4- and 8-cell embryos developed to morula were 96.7, 95.6 and 96.9%, respectively, and embryos developed to blastocysts were 88.5, 89.7 and 90.8%, respectively. 4. Conspicuous differences in size of volume and inner cell masses between single and double blastocysts were observed. Although a single blastocolic cavity was formed in most double blastocysts, several formed two distinct cavities from the very beginning.

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Inter-Cell Cooperative Scheduling with Carrier Aggregation in LTE-Advanced System (LTE-Advanced 시스템의 반송파 집성(Carrier Aggregation)을 고려한 셀간 협력 스케쥴링 기법)

  • Yang, Chan S.;Cho, Kumin;Yu, Takki;Kang, Chung G.
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.39B no.3
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    • pp.151-161
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    • 2014
  • 3GPP LTE-Advanced (Release 10) system specifies carrier aggregation (CA) to enable high data rate on using multiple frequency bands, including the variout CA-specific deployment scenarios. Considering one of those scenarios in which the different directional sector antenna is employed by each frequency band, we propose a per-carrier cell selection scheme that can improve the average throughput of the cell-edge users by allowing each user equipment (UE) to select the frequency band of the adjacent cell. Furthermore, a distributed algorithm for inter-cell copperative scheduling in this scheme is proposed to support proportional fairness among the cells. It has been shown that the proposed scheduling algorithm for the per-carrier cell selection scheme improves the cell-edge user throughput roughly by 50% over that of the conventional scheme.

Molecular association of CD98, CD29, and CD147 critically mediates monocytic U937 cell adhesion

  • Kim, Mi-Yeon;Cho, Jae Youl
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.5
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    • pp.515-523
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    • 2016
  • Adhesion events of monocytes represent an important step in inflammatory responses induced by chemokines. The ${\beta}1$-integrin CD29 is a major adhesion molecule regulating leukocyte migration and extravasation. Although several adhesion molecules have been known as regulators of CD29, the molecular interactions between CD29 and its regulatory adhesion molecules (such as CD98 and CD147) have not been fully elucidated. Therefore, in this study, we examined whether these molecules are functionally, biochemically, and cell-biologically associated using monocytic U937 cells treated with aggregation-stimulating and blocking antibodies, as well as enzyme inhibitors. The surface levels of CD29, CD98, and CD147 (but not CD43, CD44, and CD82) were increased. The activation of CD29, CD98, and CD147 by ligation of them with aggregation-activating antibodies triggered the induction of cell-cell adhesion, and sensitivity to various enzyme inhibitors and aggregation-blocking antibodies was similar for CD29-, CD98-, and CD147-induced U937 cell aggregation. Molecular association between these molecules and the actin cytoskeleton was confirmed by confocal microscopy and immunoprecipitation. These results strongly suggest that CD29 might be modulated by its biochemical and cellular regulators, including CD98 and CD147, via the actin cytoskeleton.

Optical Detection of Red Blood Cell Aggregation under vibration (진동장에서의 적혈구 응집성을 측정하는 광학적 방법)

  • Jang, Ju-Hee;Park, Myung-Su;Ku, Yun-Hee;Shin, Se-Hyun
    • Proceedings of the KSME Conference
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    • 2004.11a
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    • pp.1510-1515
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    • 2004
  • Aggregability of red blood cells (RBCs) was determined by a laser backscattering light analysis in a microfluidic channel. Available techniques for RBC aggregation often adopt a rotational Couette-flow using bob-and-cup system for disaggregating RBCs, which causes the system to be complex and expensive. A disposable microfluidic channel and vibration generating mechanism were used in the proposed new detection system for RBC aggregation. Prior to measurement, RBC aggregates in a blood sample were completely disaggregated by applying vibration-induced shear. With the present apparatus, the aggregation indexes of RBCs can be easily measured with small quantities of blood sample. The measurements with the present aggregometer were compared with those of LORCA and showed a strong correlation between them. The aggregability of the defibrinogenated blood RBCs is markedly lower than that of the normal RBCs. The noble feature of this design is the vibration-induced disaggregation mechanism, which enables to incorporate disposable element that holds the blood sample.

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