• Title/Summary/Keyword: Cell manipulation

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Controlling the Gene Expression of Corynebacterium diphtheria Toxin-A Using the Tet-On System in Mouse Embryonic Stem Cells. (Mouse Embryonic Stem Cell에서 Tetracycline-Inducible System(Tet-on System)을 이용한 Corynebacterium diphtheria Toxin-A유전자의 발현 조절)

  • 박재균;임수빈;송지환
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.11-15
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    • 2004
  • Embryonic stem (ES) cells are derived from the inner cell mass of the blastocyst-stage embryos that can be propagated indefinitely and, at the same time, can be differentiated into all the cell types that constitute the body. Current research using ES cells is mainly focused on the efficient generation of specific cell types by employing optimal differentiation conditions, which often requires the genetic manipulation of ES cells. As a way of developing an efficient system to regulate foreign gene expression in ES cells, we have inserted the gene encoding Corynebacterium diphtheria toxin-A (DTA) into an autonomously induced plasmid under positive doxycycline control ('Tet-on' system). In this study, we demonstrate that this system can lead to the cell death of mouse ES cells by the induction of DTA expression when exposed to the tetracycline derivative, doxycycline. MTT assay showed that this induction resulted in the apoptosis of ES cells.

Cellular Force Sensing for Force Feedback-Based Biological Cell Injection (힘 피드백 기반의 세포조작을 위한 세포막 침습력 측정)

  • Kim, Deok-Ho;Yun, Seok;Kang, Hyun-Jae;Kim, Byung-Kyu
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.27 no.12
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    • pp.2079-2084
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    • 2003
  • In biological cell manipulation, manual thrust or penetration of an injection pipette into an embryo cell is currently performed by a skilled operator, relying on visual feedback information only. Accurately measuring cellular forces is a requirement for minimally invasive cell injections. Moreover, the cellular force sensing is essential in investigating the biophysical properties for cell injury and membrane modeling studies. This paper presents cellular force measurements for the force feedback-based biomanipulation. Cellular force measurement system using piezoelectric polymer sensor is implemented to measure the penetration force of a zebrafish egg cell. First, measurement system setup and calibration are described. Second, the force feedback-based biomanipulation is experimentally carried out. Experimental results show that it successfully supplies real-time cellular force feedback to the operator at tens of uN and thus plays a main role in improving the reliability of biological cell injection tasks.

Effects of Autotransfusion Using Cell Saver in Cardiovascular Surgery (심혈관수술에서 cell saver를 이용한 자가수혈)

  • 구자홍
    • Journal of Chest Surgery
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    • v.26 no.4
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    • pp.255-259
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    • 1993
  • The efficacy of the autotransfusion system is a reducing the need of intraoperative and postoperative transfusion in cardiovascular surgery. Between January 1990 and December 1991, we experienced 23 cases of autotransfusion using Haemonetic Cell Saver in cardiovascular surgery [Experimental group]. Another 13 cases which were taken similiar operations without Cell Saver during same period [Control group]. The amounts of blood transfused are 4.23 1.84 units in Control group, 2.82 1.84 units in Experimental group. Postoperatively, both groups showed decreased platelet counts, mild prolongation of prothrombin and partial thromboplastin time compred to preoperative value [P<0.001], but there were no significant differences between two group [P=NS]. Plasma hemoglobin was markedly increased in Experimental group compared with Control group [p<0.05]. In Experimental group, amount of average processed blood by Cell Saver was 700ml of which composition was hemoglobin 17mg/dl, hematocrit 50.0%, RBC 5,590,000/ml, WBC 7500/ml, and platelet 40,000/ml. The culture of the processed blood revealed no growth of the organisms. Conclusively, Cell Saver autotransfusion system is a simple, safe, and cost effective method especially in the cases associated with massive bleeding. However, it requires familiarity with system, gentle manipulation of suction tip, and careful selection of candidates to obtain maximal benefits .

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Cellular Force Measurement for Force Feedback-Based Biomanipulation (힘반향 기반의 바이오매니퓰레이션을 위한 세포 조작력 측정)

  • Kim, Duk-Ho;Kim, Byung-Kyu;Yoon, Seok;Kang, Hyun-Jae
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 2003.06a
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    • pp.237-240
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    • 2003
  • In biological cell manipulation, manual thrust or penetration of an injection pipette into an embryo cell is currently performed by a skilled operator, relying on visual feedback information only. Accurately measuring cellular forces is a requirement for minimally invasive cell injections. Moreover, the cellular farce sensing is essential in investigating the biophysical properties for cell injury and membrane modeling studies. This paper presents cellular force measurements for the force feedback-based biomanipulation. Cellular force measurement system using piezoelectric polymer sensor is implemented to measure the penetration force of a zebrafish egg cell. First, measurement system setup and calibration are described. Second, the force feedback-based biomanipulation is experimentally carried out. Experimental results show that it successfully supplies real-time cellular force feedback to the operator at several tens of uN and thus plays a main role in improving the reliability of biological cell injection tasks.

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DESIGN OF A CHECK VALVE FOR FEEDING BIOLOGICAL CELLS ONE BY ONE (세포의 개별 공급을 위한 체크 밸브의 설계)

  • Choi, Jung-Hoon;Lee, Sang-Wook;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 1994.07a
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    • pp.85-87
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    • 1994
  • Feeding biological cells one by one is the key point in the manipulation of cells. The conventional valve systems have many difficulties in feeding cells one by one, because they shut the whole flow of fluids when they are closed and have possibilities of breaking the fragile cells. They need some other equipments for continuous supply of suspension and to protect the cells. We design a check valve for feeding biological cells one by one using polyimide all the silicon substrate. The cells are fed by hydraulic pressure through the isotropically etched cavity. When the suspension flows continuously along the channel the valve is bent by hydraulic pressure and a cell is fed to the outlet. We have studied a cell fusion device fabricated with polyimide and electroplating. If the designed check valve is located in front of the cell fusion device it is helpful to fuse two different kinds of cells.

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Recent Advances in Cell Therapeutics for Systemic Autoimmune Diseases

  • Youngjae Park;Seung-Ki Kwok
    • IMMUNE NETWORK
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    • v.22 no.1
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    • pp.10.1-10.17
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    • 2022
  • Systemic autoimmune diseases arise from loss of self-tolerance and immune homeostasis between effector and regulator functions. There are many therapeutic modalities for autoimmune diseases ranging from conventional disease-modifying anti-rheumatic drugs and immunosuppressants exerting nonspecific immune suppression to targeted agents including biologic agents and small molecule inhibitors aiming at specific cytokines and intracellular signal pathways. However, such current therapeutic strategies can rarely induce recovery of immune tolerance in autoimmune disease patients. To overcome limitations of conventional treatment modalities, novel approaches using specific cell populations with immune-regulatory properties have been attempted to attenuate autoimmunity. Recently progressed biotechnologies enable sufficient in vitro expansion and proper manipulation of such 'tolerogenic' cell populations to be considered for clinical application. We introduce 3 representative cell types with immunosuppressive features, including mesenchymal stromal cells, Tregs, and myeloid-derived suppressor cells. Their cellular definitions, characteristics, mechanisms of immune regulation, and recent data about preclinical and clinical studies in systemic autoimmune diseases are reviewed here. Challenges and limitations of each cell therapy are also addressed.

The Role of PI3K/AKT Pathway and NADPH Oxidase 4 in Host ROS Manipulation by Toxoplasma gondii

  • Choi, Hei Gwon;Gao, Fei-Fei;Zhou, Wei;Sun, Pu-Reum;Yuk, Jae-Min;Lee, Young-Ha;Cha, Guang-Ho
    • Parasites, Hosts and Diseases
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    • v.58 no.3
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    • pp.237-247
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    • 2020
  • Dendritic cell is one of the first innate immune cell to encounter T. gondii after the parasite crosses the host intestinal epithelium. T. gondii requires intact DC as a carrier to infiltrate into host central nervous system (CNS) without being detected or eliminated by host defense system. The mechanism by which T. gondii avoids innate immune defense of host cell, especially in the dendritic cell is unknown. Therefore, we examined the role of host PI3K/AKT signaling pathway activation by T. gondii in dendritic cell. T. gondii infection or T. gondii excretory/secretory antigen (TgESA) treatment to the murine dendritic cell line DC2.4 induced AKT phosphorylation, and treatment of PI3K inhibitors effectively suppressed the T. gondii proliferation but had no effect on infection rate or invasion rate. Furthermore, it is found that T. gondii or TgESA can reduce H2O2-induced intracellular reactive oxygen species (ROS) as well as host endogenous ROS via PI3K/AKT pathway activation. While searching for the main source of the ROS, we found that NADPH oxidase 4 (NOX4) expression was controlled by T. gondii infection or TgESA treatment, which is in correlation with previous observation of the ROS reduction by identical treatments. These findings suggest that the manipulation of the host PI3K/AKT signaling pathway and NOX4 expression is an essential mechanism for the down-regulation of ROS, and therefore, for the survival and the proliferation of T. gondii.

Influence of Chromosome Number on Cell Growth and Cell Aging in Yeast (효모에서 염색체의 수가 세포성장과 노화에 미치는 영향)

  • Kim, Yeon-Hee
    • Journal of Life Science
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    • v.26 no.6
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    • pp.646-650
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    • 2016
  • The influence of chromosome number on cell growth and cell aging was investigated in various yeast strains that have many artificial chromosomes constructed using a chromosome manipulation technique. Host strain FY833 and the YKY18, YKY18R, YKY24, and YKY30 strains harboring 16 natural chromosomes, 18 chromosomes, 18 chromosomes containing rDNA chromosome, 24 chromosomes, and 30 chromosomes, respectively, were used, and the specific growth rate of each strain was compared. The specific growth rates in the YKY18 and YKY24 strains were indistinguishable from that in the host strain, while those of the YKY18R and YKY30 strains were reduced to approximately 25% and 40% of the host strain level, respectively. Subsequently, the replicative life span was examined to investigate the relationship between the number of chromosomes and cell aging, and the life span was decreased to approximately 14% and 45% of the host strain level in the YKY24 and YKY30 strains, respectively. Moreover, telomere length, well known as a senescence factor, was shorter and more diversified in the strain, showing decreased life span. Therefore, these results suggest the possibility that an increase in the number of chromosomes containing artificial chromosomes caused cell aging, and we expected these observations would be applied to improve industrial strain harboring of versatile and special artificial chromosomes.

Seaweed Biotechnology and Biologically Active Substances

  • Hong, Yong-Ki
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.2
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    • pp.59-62
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    • 2006
  • Seaweed biotechnology is a multidisciplinary subject to produce food, pharmaceuticals, chemicals, and environmental remediation materials from seaweed resources. It uses various techniques of cell culture, enzyme reaction and genetic manipulation to increase the production efficiency of useful seaweeds or their products. Firstly, an overview of key topics will be introduced in the fields of seaweed tissue culture, strain improvement, genetic analysis briefly as basic techniques. Secondly, some biologically active substances such as anti-inflammatory and antifouling substances that have been screened in my laboratory will be focused.

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Plant Defence Responses: Current Status and Future Exploitation

  • Yun, Byung-Wook;Gray J, Loake
    • Journal of Plant Biotechnology
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    • v.4 no.1
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    • pp.1-6
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    • 2002
  • Plants have developed a sophisticated battery of defence responses to protect themselves against attempted pathogen ingress. Manipulation of these defence mechanisms may provide significant opportunities for crop improvement. While plant resistance genes have had a long service history in plant breeding, they possess significant limitations. Recent advances are now providing significant insights into strategies designed to increase the field durability of this class of genes. Hypersensitive cell death is a common feature underlying the deployment of plant defence responses against biographic pathogens. In contrast, necrotrophic pathogens actively kill plant cells. Recently, transgenic plants have been developed that either promote or suppress cell death, providing resistance against either biotrophic or necrotrophic pathogens respectively. Methyl-jasmonate is a key signalling molecule in the establishment of resistance against some fungal pathogens. Increasing the concentration of this molecule in plant cells has been shown to increase resistance against Botrytis cineria, without significantly imparting plant growth or development. Due to the multifarious infection strategies employed by plant pathogens, how-ever, it is unlikely a single commercial product will prove a panacea for global disease control. Future stategies will more likely entail an integrated disease management approach.