• Title/Summary/Keyword: cell culture

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Design, Fabrication, and Application of a Microfluidic Device for Investigating Physical Stress-Induced Behavior in Yeast and Microalgae

  • Oh, Soojung;Kim, Jangho;Ryu, Hyun Ryul;Lim, Ki-Taek;Chung, Jong Hoon;Jeon, Noo Li
    • Journal of Biosystems Engineering
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    • v.39 no.3
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    • pp.244-252
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    • 2014
  • Purpose: The development of an efficient in vitro cell culture device to process various cells would represent a major milestone in biological science and engineering. However, the current conventional macro-scale in vitro cell culture platforms are limited in their capacity for detailed analysis and determination of cellular behavior in complex environments. This paper describes a microfluidic-based culture device that allows accurate control of parameters of physical cues such as pressure. Methods: A microfluidic device, as a model microbioreactor, was designed and fabricated to culture Saccharomyces cerevisiae and Chlamydomonas reinhardtii under various conditions of physical pressure stimulus. This device was compatible with live-cell imaging and allowed quantitative analysis of physical cue-induced behavior in yeast and microalgae. Results: A simple microfluidic-based in vitro cell culture device containing a cell culture channel and an air channel was developed to investigate physical pressure stress-induced behavior in yeasts and microalgae. The shapes of Saccharomyces cerevisiae and Chlamydomonas reinhardtii could be controlled under compressive stress. The lipid production by Chlamydomonas reinhardtii was significantly enhanced by compressive stress in the microfluidic device when compared to cells cultured without compressive stress. Conclusions: This microfluidic-based in vitro cell culture device can be used as a tool for quantitative analysis of cellular behavior under complex physical and chemical conditions.

Effects of Chitosan on Human Gingival Fibroblasts in Vitro (키토산이 치은섬유아세포에 미치는 영향)

  • Kim, Ok-Su;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.32 no.1
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    • pp.235-247
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    • 2002
  • The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of human gingival fibroblasts. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells were inoculated in the multiwell plates coated with chitosan in concentration of 0.02, 0.2, and 2 mg/ml. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized nodules was evaluated after 21 days of culture. The results were as follows : The morphology of cells on the chitosan-coated well was round or spheric. Round cells were aggregated since 6 hours of culture and showed nodule-like appearance after 24 hours of culture and did not achieved confluency at 7 days. The attachment of gingival fibroblasts was inhibited by chitosan coating with a tendency of dose dependent pattern. But, cellular activity of unit cell was higher than control. The proliferation of gingival fibroblasts was inhibited by chitosan coating at 2 mg/ml(P<0.01), while the cell proliferation at 0.02, 0.2 $mg/m{\ell}$ was comparable to the control well. Total alkaline phosphatase activity was inhibited by chitosan coating and decreased in the course of time. While ALP activity of unit cell was the highest at 2mg/ml after 4 days of culture. Finally, gingival fibroblasts produced the mineralized nodule at 2 mg/ml. In summary, the attachment, proliferation, and alkaline phosphatase activity of gingival fibroblasts were influenced differently by the concentration of coated chitosan. From this study, it could be used as the matrix of tissue engineering for gingiva without inhibition on proliferation of gingival fibroblasts using chitosan at the optimal concentration (0.02mg/ml).

Effects of in vitro Culture Period of Reconstructed Embryos and Genetic Background of Feeder Cells on Establishment of Embryonic Stem Cells Derived from Somatic Cell Nuclear Transfer Blastocysts in Pigs

  • Han, Na Rae;Baek, Song;Lee, Yongjin;Lee, Joohyeong;Yun, Jung Im;Lee, Eunsong;Lee, Seung Tae
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.1
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    • pp.86-93
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    • 2020
  • The establishment of porcine embryonic stem cells (ESCs) from porcine somatic cell nuclear transfer (SCNT) blastocysts is influenced by in vitro culture day of porcine reconstructed embryo and feeder cell type. Therefore, the objective of the present study was to determine the optimal in vitro culture period for reconstructed porcine SCNT embryos and mouse embryonic fibroblast (MEF) feeder cell type for enhancing colony formation efficiency from the inner cell mass (ICM) of porcine SCNT blastocysts and their outgrowth. As the results, porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days showed significantly increased efficiency in the formation of colonies, compared to those for 7 days. Moreover, MEF feeder cells derived from outbred ICR mice showed numerically the highest efficiency of colony formation in blastocysts produced through in vitro culture of porcine SCNT embryos for 8 days and porcine ESCs with typical ESC morphology were maintained more successfully over Passage 2 on outbred ICR mice-derived MEF feeder cells than on MEF feeder cells derived from inbred C57BL/6 and hybrid B6CBAF1 mice. Overall, the harmonization of porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days and MEF feeder cells derived from outbred ICR mice will greatly contribute to the successful establishment of ESCs derived from porcine SCNT blastocysts.

Production of Emulsan by Acinetobacter calcoaceticus RAG-1 under Various Culture Modes (여러 배양방법하에서 Acinetobacter calcoaceticus RAG-1에 의한 Emulsan의 생산)

  • 강병철;이필경장호남
    • KSBB Journal
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    • v.6 no.4
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    • pp.389-394
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    • 1991
  • Emulsan is an extracellular emulsifying agent produced by the hydrocarbon-degrading Acinetobacter species RAG-1. In this study emulsan production of Acinetobacter calcpaceticus RAG-1 was investigated under various culture modes such as batch, fed-batch, membrane cell recycle, and continuous culture. The productions of emulsan under both ethanol-sufficient fed-batch and membrane cell recycle cultures were all 15.0U/ml, which was 53% increase in emulsan activity compared to that of pH controlled batch culture. Emulsan production was found to be strongly dependent on the residual ethanol concentration. In continuous culture the emulsan productivity increased with dilution rate.

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Selection of Cell Source and the Effect of pH and MS Macronutrients on Biomass Production in Cell Cultures of Tongkat Ali (Eurycoma longifolia Jack)

  • Siregar Luthfi Aziz Mahmud;Chan Lai-Keng;Boey Peng-Lim
    • Journal of Plant Biotechnology
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    • v.5 no.2
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    • pp.131-135
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    • 2003
  • Callus and cell suspension cultures of Eurycoma longifolia Jack were initiated from leaves of different trees. The leaf explant of tree Eu9 produced the most calli and also induced high cell biomass in the cell suspension culture. Optimum production of cell biomass could be initiated in proliferating culture medium with a pH of 5.75 prior to autoclaving. The effects of macronutrient inorganic salts of Murashige and Skoog (MS) liquid medium supplemented with X on production of cell biomass of Eurycoma longifolia were also investigated. The highest cell biomass was produced in MS medium containing macronutrients of $21\;mM\;NH_4NO_3,\;12.25\;mM\;KNO_3,\;3.00\;mM\;CaCl_2.2H_2O,\;0.575\;mM\;MgSO_4.7H_2O$, and $1.83\;mM\;KH_2PO_4$. A new medium labeled as TAM was formulated for the production of Eurycoma longifolia cell biomass in the cell suspension culture.

Production of 10-deacetylbaccatin III in Taxus cuspidata Suspension Cell Culture (주목 현탁세포배양을 이용한 10-deacetylbaccatin III 생산)

  • Lee, Gue-Wha;Kim, Dong-Il
    • KSBB Journal
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    • v.14 no.6
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    • pp.665-671
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    • 1999
  • In this study, enhanced production of 10-deacetylbaccatin III(10-DAB), a precursor of taxol in semisynthesis, was investigated in Taxus cuspidata cell suspension cultures. The effects of initial inoculum size and sugar concentration were examined to prove the relationship between the production of 10-DAB and cell growth. The cell growth was found to be stimulated in Schenk and Hildebrandt(SH) medium. The lower the inoculum size as well as initial sugar concentration, the faster the cell growth rate. When the initial sugar concentration was dept low, the production of 10-DAB into medium was increased. By using perfusion culture, continuous cell growth was possible until the end of culture and more than 34.67 g/L of cell concentration could by obtained. This is about 2.5 times higher level than that of control batch culture.

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A study on the Effects of Organism Vigor Information Solution to the Growth of Tumor-Derived Animal Cell (생물활성정보수가 종양 유래 동물세포 성장에 미치는 영향에 관한 고찰)

  • Jang, M.Y.;Lee, K.S.;Shin, J.W.;Mun, C.W.
    • Journal of Biomedical Engineering Research
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    • v.32 no.3
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    • pp.185-190
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    • 2011
  • The purpose of this study is to investigate the influence of organism vigor information solution(OVIS) on the animal cell growth and to set up a condition for cell culture. We investigated the reactions on the MG-63 and MCF-7 cell line in mixture culture media with various amount of REVIEW solution, which is an example among various OVIS. DMEM-HG was used as basic media. The concentration range of the mixture was limited from 0% to 15%. MTT assay is used for cell viability test. The cell was incubated for 14days and the MTT assay was performed on day 1, 3, 7, 10 and 14 throughout the experiment. We used the ELISA reader to measure the Optical Density(O.D) at 595 nm wavelength filter. MCF-7 was linearly proliferated according to culture time and concentrations of OVIS. On the other hand, MG-63 cells were measured the highest O.D value at 12%. The growth rates of both cells cultured in mixed culture media with OVIS are much higher than those in only basic media, DMEMHG, after 14days. It was confirmed that cell cultured at OVIS grows rapidly at certain period although cells showed a negative effect in initial stage.

Physiological studies on cell division by the technique of synchronous culture of chlorella (I) (클로렐라의 동조배양법에 의한 세포분열의 생리학적 연구 1)

  • 이영녹;이종삼
    • Korean Journal of Microbiology
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    • v.7 no.1
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    • pp.1-9
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    • 1969
  • Changes in the phosphorylation of Chlorella cells during the life cycle the aulotrophic and micotrophic synchronous culture were followed under the light and dark. 1. In the autotrophic culture of Chlorella the amounts of esterified phosphate compounds of the algal cell under the light increased during the growing period and decreased strikingly in the ripening period showing a peak at the $L_1$ i/-cell stage. 2. TRhe amount of total esterified phosphate compounds of the cell under the dark, however, decreased during the growing period and then kept fairly constnat during the ripening nad division periods showing the greates activity of the oxidative phosphorylation in the early growing stage. 3. It is presumed that the energy requirement of the dividing algal cell in the autotrophic culture is fulfilled prior to the nuclear division mostly by the photosynthetic phosphorylation. 4. In the mixotrophic culture, the amount of esterified phosphate compounds of the algal cells under the light increased during the growing period and decreased during the late ripening and early division periods showing a peak in the $L_2$-cell stage as in the case of the phosphorylation under the dark. 5. The phosphorylation of the fell grown in the glucose medium is more active under the dark than under the light in the stages of the growing and early ripening periods. 6. It is considered that the excess glucose in the algal cell not only promotes the oxidative phosphorylation but also inhibits the photosynthetic phosphorylation of the cell. 7. It is presumed that the energy requirement of the dividing algal cell in the glucose medium is fulfilled prior to the nuclear division by the combined action of oxidative and photosynthetic phosphorylation, mostly by the oxidative phosphorylation.

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Effect of Culture Medium Containing Chicken Serum on Growth and Differentiation of QM7 Quail Muscle Cells (닭 혈청을 포함한 배양액 조성 변화가 QM7 메추리 근육세포의 성장 및 분화에 미치는 영향 분석)

  • Choi, Sarang;Lee, Sang In;Shin, Sangsu
    • Korean Journal of Poultry Science
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    • v.49 no.2
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    • pp.109-114
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    • 2022
  • QM7 cell, a quail muscle cell line, has been used in various studies. In cell culture, it is well known that the culture medium has a significant influence on cell growth and maintenance of cell characteristics. This study aimed to adjust the culture medium to make it more suitable for QM7 muscle cells. A culture medium was prepared by adding 2% chicken serum (CS) instead of 10% tryptose phosphate broth (TPB) to the conventional culture medium. In the culture medium prepared with CS, the QM7 muscle cells changed from a pointed, thin to a broader shape. In addition, they grew and divided more rapidly in the new culture medium than in the conventional culture medium. The number of cells increased faster in the CS-containing culture medium from day two after passaging, and significantly increased from day three. The muscle cells grown in the medium containing CS maintained their undifferentiated state prior to differentiation. Myotubes formed well when the cells were maintained in CS-containing medium, resulting in a longer length and uniformity. According to the above results, it is clear that the culture of QM7 myocytes using a medium containing CS rather than TPB helps obtain better results in cell maintenance and differentiation.

Characterization of Dye Decolorization in Cell-Free Culture Broth of Trametes versicolor CBR43

  • Ryu, Hyun;Ryu, Hee Wook;Cho, Kyung-Suk
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.155-160
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    • 2017
  • The dye decolorization rate in a cell-free culture broth of the white-rot fungus Trametes versicolor CBR43 was studied, including the effects of inhibitors of NaCl, Zn(II), and Cd(II) on dye decolorization activity. The maximum rates of dye decolorization in cell-free culture broth were 1,410, 44.7, 41.2, and $0.19{\mu}mol{\cdot}l^{-1}{\cdot}min^{-1}$ for Acid Blue 62, Acid Black 175, Reactive Blue 4, and Acid Red 114, respectively. The inhibition effects of NaCl, Zn(II), and Cd(II) on dye decolorization were quantitatively compared using the half maximal inhibition concentration ($IC_{50}$), which indicates the concentration of an inhibitor required for 50% inhibition. Based on $IC_{50}$ values, dye decolorization in the cell-free culture broth of CBR43 was most potently inhibited by Cd(II), whereas the inhibitory effect of NaCl was relatively low. The dye decolorization rates and $IC_{50}$ data can be used in the design and development of a dye-wastewater treatment process using T. versicolor CBR43 and its operating factors.