• Title/Summary/Keyword: Flow-through Cell

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Optimization and Mathematical Modeling of the Transtubular Bioreactor for the Production of Monoclonal Antibodies from a Hybridoma Cell Line

  • Halberstadt, Craig R.;Palsson, Bernhanrd O.;Midgley, A.Rees;Curl, Rane L.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.3
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    • pp.163-170
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    • 2002
  • This report describes the use of a transtubular bioreactor to study the relative effects of diffusion versus perfusion of medium on antibody production by a hybridoma cell line. The study was performed with a high-density cell culture maintained in a serum-free, low-protein medium for 77 days. It was determined that the reactor possessed a macro-mixing pattern residence time distribution similar to a continuous stirred tank reactor (CSTR), However, due to the arrangement of the medium lines in the reactor, the flow patterns for nutrient distribution consist of largely independent medium path lengths ranging from short to long. When operated with cyclic, reversing, transtubular medium flow, some regions of the reactor (with short residence times) are more accessible to medium than others (with long residence times). From this standpoint, the reactor can be divided into three regions: a captive volume, which consists of medium primarily delivered via diffusion; a lapped volume, which provides nutrients through unilateral convection; and a swept volume, which operates through bilateral convection. The relative sizes of these three volumes were modified experimentally by changing the period over which the direction of medium flow was reversed from 15 min (larger captive volume) to 9 h (larger swept volume). The results suggest that antibody concentration increases as the size of the diffusion-limited (captive) volume is increased to a maximum at around 30 min with a sharp decrease thereafter. As reflected by changes in measured consumption of glucose and production of lactate, no significant difference in cellular metabolism occurred as the reactor was moved between these different states. These results indicate that the mode of operation of the transtubular bioreactor may influence antibody productivity under serum-free, low-protein conditions with minimal effects on cellular metabolism.

LINC01272 Suppressed Cell Multiplication and Induced Apoptosis Via Regulating MiR-7-5p/CRLS1 Axis in Lung Cancer

  • Ma, Xuan;Liu, Yang;Tian, Hao;Zhang, Bo;Wang, Meiling;Gao, Xia
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.921-932
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    • 2021
  • LINC01272 is a long non-coding RNA (lncRNA) that has been considered as a biomarker for many diseases including lung squamous cell carcinoma. Here, we investigated the function and mechanism of LINC01272 on lung cancer (LC). The differential expression of LINC01272 in LC and normal samples was analyzed by GEPIA based on the data from TCGA-LUAD database, as survival prognosis was analyzed through Kaplan-Meier Plotter. LINC01272 overexpression plasmid and miR-7-5p mimic were transfected into A549 and PC-9 cells. LINC01272, miR-7-5p and cardiolipin synthase 1 (CRLS1) mRNA expression was measured by quantitative reverse transcription-polymerase chain reaction. Cell viability was detected through MTT assay. Cell multiplication was evaluated by cell formation assay. Cell apoptosis was assessed through flow cytometry assay. Through bioinformatics, the target miRNA of LINC01272 and downstream genes of miR-7-5p were predicted. The targeting relationship was tested by dual luciferase reporter analysis. CRLS1, B-cell lymphoma-2 (Bcl-2), BCL2-associated X (Bax) and cleaved caspase-3 protein levels were detected through western blot. LINC01272 was downregulated in LC and low LINC01272 expression had poor prognosis. In A549 and PC-9 cells, LINC01272 inhibited cell viability and multiplication and induced apoptosis. LINC01272 negatively regulated miR-7-5p and CRLS1 was a target of miR-7-5p. MiR-7-5p reversed the effect of LINC01272 on viability, multiplication, apoptosis and expression of miR-7-5p and CRLS1 as well as apoptosis-related factors (Bcl-2, Bax and cleaved caspase-3). LINC01272 suppressed cell multiplication and induced apoptosis via regulating the miR-7-5p/CRLS1 axis in LC.

Design of Serpentine Flow-field Stimulating Under-rib Convection for Improving the Water Discharge Performance in Polymer Electrolyte fuel cells (고분자전해질 연료전지의 물 배출 성능 향상을 위한 촉매층 공급 대류 촉진 사행성 유동장 설계)

  • Choi, Kap-Seung;Bae, Byeong-Cheol;Park, Ki-Won;Kim, Hyung-Man
    • Journal of the Korean Electrochemical Society
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    • v.15 no.2
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    • pp.74-82
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    • 2012
  • Proton exchange membrane fuel cell performance is changed by the complicated physical phenomenon. In this study, water discharge performance of proton exchange membrane fuel cell were performed numerically to compare serpentine channel flow fields of 5-pass 4-turn serpentine and 25 $cm^2$ reaction surface between with and without sub-channel at the rib. Through the supplement of sub channel flow field, it is shown from the results that water removal characteristic inside channel improves because the flow direction of under-rib convection is changed into the sub channel. Reacting gases supplied from entrance disperse into sub channel flow field and electrochemical reaction occurs uniformly over the reaction surface. The results obtained that total current density distributions become uniform because residence time of reacting gases traveling to sub-channel flow field is longer than to main channel.

Spherical and cylindrical microencapsulation of living cells using microfluidic devices

  • Hong, Joung-Sook;Shin, Su-Jung;Lee, Sang-Hoon;Wong, Edeline;Cooper-White, Justin
    • Korea-Australia Rheology Journal
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    • v.19 no.3
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    • pp.157-164
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    • 2007
  • Microencapsulation of cells within microfluidic devices enables explicit control of the membrane thickness or cell density, resulting in improved viability of the transplanted cells within an aggressive immune system. In this study, living cells (3T3 and L929 fibroblast cells) are encapsulated within a semi-permeable membrane (calcium crosslinked alginate gel) in two different device designs, a flow focusing and a core-annular flow focusing geometry. These two device designs produce a bead and a long microfibre, respectively. For the alginate bead, an alginate aqueous solution incorporating cells flows through a flow focusing channel and an alginate droplet is formed from the balance of interfacial forces and viscous drag forces resulting from the continuous (oil) phase flowing past the alginate solution. It immediately reacts with an adjacent $CaCl_2$ drop that is extruded into the main flow channel by another flow focusing channel downstream of the site of alginate drop creation. Depending on the flow conditions, monodisperse microbeads of sizes ranging from $50-200\;{\mu}m$ can be produced. In the case of the microfibre, the alginate solution with cells is extruded into a continuous phase of $CaCl_2$ solution. The diameter of alginate fibres produced via this technique can be tightly controlled by changing both flow rates. Cell viability in both forms of alginate encapsulant was confirmed by a LIVE/DEAD cell assay for periods of up to 24 hours post encapsulation.

Inhibitory Effect of Lemon Oil on Apoptosis in Astrocytes (신경교(神經膠) 성상세포(星狀細胞)에서 레몬오일에 의한 세포자멸사(細胞自滅死)의 억제효과(抑制效果))

  • Kim, Jun-Han;Kim, Tae-Heon
    • Journal of Oriental Neuropsychiatry
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    • v.11 no.1
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    • pp.37-46
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    • 2000
  • We investigated the effects of lemon pure essential oils on the heat shock-induced apoptosis in human astrocyte cell line CCF-STTG1. In previous studies, hear shock has been reported to induce the apoptosis or programmed cell death through the activation of caspase-3. Treatment of CCF-STTG1 cells with heat shock markedly induced apoptotic cell death as determined by flow cytometry. Interestingly, pretreatment of CCF-STTG1 cells with lemon pure essential oils inhibited the heat shock-induced apoptosis. Lemon also inhibited the heat shock-induced apoptosis in primary cultured rat astrocytes. To determine whether lemon inhibits the heat shock-induced activation of these apoptotic proteases, activation of CPP32 was assessed by Western blotting. Consistent with flow cytometry, DNA fragmentation and giemsa staining, heat shock-induced activation of CPP32 was blocked by lemon pure essential oil. PARP, cysteine protease substrates were fragmented as a consequence of apoptosis by heat shock. Lemon oil inhibited the PARP fragmentation. This essential oil also inhibited the heat shock-induced activation of caspase-3. These results suggest that lemon pure essential oils may modulate the apoptosis through the activation of the ICE-like caspases.

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Development of Automated Diffusion Cell for Determining In Vitro Drug Release from Transdermal Device (경피흡수제형의 in vitro 약물방출실험을 위한 연속확산 장치의 개발)

  • Byun, Young-Rho;Choi, Young-Kweon;Jeong, Seo-Young;Kim, Young-Ha
    • YAKHAK HOEJI
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    • v.34 no.3
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    • pp.161-165
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    • 1990
  • An automated, simple, and reliable method was developed for determining in vitro drug release rate from transdermal delivery dosage forms. The patch is held in position in the heating block by sandwiching it between the middle plate and the bottom plate of diffusion cell. The dissolution profile of the commercially available transdermal scopolamine patch was determined over a 72-h period, and the results were compared with those obtained with other methods; paddle-over-disk method, reciprocating method, and diffusion cell method. It was demonstrated that the flow-through method is equivalent in terms of release rate profile and accumulated released drug amount over the lifetime of the dosage form tested. Also this method is simple, reliable and reproducible. Therefore, this technique can be used in a quality control for assuring product uniformity.

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A Study on the Configuration of BOP and Implementation of BMS Function for VRFB (VRFB를 위한 BOP 구성 및 BMS 기능구현에 관한 연구)

  • Choi, Jung-Sik;Oh, Seung-Yeol;Chung, Dong-Hwa;Park, Byung-Chul
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.28 no.12
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    • pp.74-83
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    • 2014
  • This paper proposes a study on the configuration of balancing of plant(BOP) and implementation of battery management system(BMS) functions for vanadium redox flow battery(VRFB) and propose a method consists of sensor and required design specifications BOP system configuration. And it proposes an method of the functions implementation and control algorithm of the BMS for flow battery. Functions of BMS include temperature control, the charge and discharge control, flow control, level control, state of charge(SOC) estimation and a battery protection through the sensor signal of BOP. Functions of BMS is implemented by the sensor signal, so it is recognized as a very important factor measurement accuracy of the data. Therefore, measuring a mechanical signal(flow rate, temperature, level) through the BOP test model, and the measuring an electrical signal(cell voltage, stack voltage and stack current) through the VRFB charge-discharge system and analyzes the precision of data in this paper. Also it shows a good charge-discharge test results by the SOC estimation algorithm of VRFB. Proposed BOP configuration and BMS functions implementation can be used as a reference indicator for VRFB system design.

NUMERICAL STUDY FOR THE FULL-SCALE ANALYSIS OF PLATE-TYPE HEAT EXCHANGER USING ONE-DIMENSIONAL FLOW NETWORK MODEL and ε-NTU METHOD (판형 열교환기 Full-scale 해석을 위한 1차원 유동 네트워크 모델 및 ε-NTU 모델의 수치적 연구)

  • Kim, Minsung;Min, June Kee;Ha, Man Yeong
    • Journal of computational fluids engineering
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    • v.19 no.1
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    • pp.47-56
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    • 2014
  • Since a typical plate heat exchanger is made up of a huge number of unitary cells, it may be impossible to predict the aero-thermal performance of the full scale heat exchanger through three-dimensional numerical simulation due to the enormous amount of computing resources and time required. In the present study, a simple flow-network model using the friction factor correlation and a thermal-network model based on the effectiveness-number of transfer units (${\varepsilon}$-NTU) method has been developed. The complicated flow pattern inside the cross-corrugated heat exchanger has been modeled into flow and thermal networks. Using this model, the heat transfer between neighboring streams can be considered, and the pressure drop and the heat transfer rate of full-scale heat exchanger matrix are calculated. In the calculation, the aero-thermal performance of each unitary cell of the heat exchanger matrix was evaluated using correlations of the Fanning friction factor f and the Nusselt number Nu, which were calculated by unitary-cell CFD model.

The effect of PEMFC stack performance at air supply condition (공기공급 조건이 스택성능에 미치는 영향)

  • Park, Chang-Kwon;Oh, Byeong-Soo
    • Transactions of the Korean hydrogen and new energy society
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    • v.19 no.3
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    • pp.232-238
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    • 2008
  • Research has been proceeded on fuel cell which is fueled by hydrogen. Polymer electrolyte membrane fuel cell (PEMFC) is promising power source due to high power density, simple construction and operation at low temperature. But it has problems such as high cost, temperature dependent performance. These problems could be solved by experiment which is useful for analysis and optimization of fuel cell performance and heat management. In this paper, when hydrogen flows constantly at the stoichiometry of ${\xi}=1.6$, the performance of the fuel cell stack was increased and the voltage difference between each cells was decreased according to the increase of air stoichiometry by 2.0, 2.5, 3.0. Therefore, the control of air flow rate in the same gas channel is important to get higher performance. Purpose of this research is to expect operation temperature, flow rate, performance and mass transportation through experiment and to help actual manufacture of PEM fuel cell stack.

Protective Effects of Verapamil against H2O2-Induced Apoptosis in Human Lens Epithelial Cells

  • Wang, Zhuo;Wang, Dan;Li, Yan;Zhang, Xiuli
    • Biomolecules & Therapeutics
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    • v.22 no.6
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    • pp.553-557
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    • 2014
  • Verapamil is used in the treatment of hypertension, angina pectoris, and atrial fibrillation. Recently, several studies have demonstrated that verapamil increased the optic nerve head blood flow and improved the retrobulbar circulation. All these show that verapamil is potentially useful for ophthalmic treatment. Thus, the aim of this study is to investigate whether verapamil could protect human lens epithelial cell (HLEC) from oxidative stress induced by $H_2O_2$ and the cellular mechanism underlying this protective function. The viability of HLEC was determined by the MTT assay and apoptotic cell death was analyzed by Hoechst 33258 staining. Moreover, Caspase-3 expression was detected by immunocytochemistry and flow cytometry analysis. We also detected Caspase-3 mRNA expression by reverse-transcription-polymerase chain reaction and the GSH content in cell culture. The results showed that oxidative stress produced significant cell apoptotic death and it was reduced by previous treatment with the verapamil. Verapamil was effective in reducing HLEC death mainly through reducing the expression level of apoptosis-related proteins, caspase-3, and increasing glutathione content. Therefore, it was suggested that verapamil was effective in reducing HLEC apoptosis induced by $H_2O_2$.