• Title/Summary/Keyword: cell production

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Increased lignan biosynthesis in the suspension cultures of Linum album by fungal extracts

  • Bahabadi, Sedigheh Esmaeilzadeh;Sharifi, Mozafar;Safaie, Naser;Murata, Jun;Yamagaki, Tohru;Satake, Honoo
    • Plant Biotechnology Reports
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    • v.5 no.4
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    • pp.367-373
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    • 2011
  • Linum album accumulates anti-tumor podophyllotoxin (PTOX) and its related lignans, which were originally isolated from an endangered species Podophyllum. In the present study, we examined the effects of five fungal extracts on the production of lignans in L. album cell cultures. Fusarium graminearum extract induced the highest increase of PTOX [$143{\mu}g\;g^{-1}$ dry weight (DW) of the L. album cell culture], while Rhizopus stolonifer extract enhanced the accumulation of lariciresinol up to $364{\mu}g\;g^{-1}$ DW, instead of PTOX. Typical elicitors, such as chitin, chitosan, or methyl jasmonate (MeJA), were shown to be less effective in lignan production in L. album cell cultures. These results verified the advantages of fungal extracts to increase lignan production in L. album cell culture, and suggested potential on-demand metabolic engineering of lignan biosynthesis using differential fungal extracts.

Experimental Effects of Taklihwangki-Tang on the Anti-Cancer And Immuno-Action (托裏黃기湯이 抗癌 및 免疫作用에 미치는 實驗的 效果)

  • Jeong, Dong-Hwan;Choi, Jung-Hwa;Kim, Jong-Han;Jeong, Woo-Hyun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.15 no.2
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    • pp.118-130
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    • 2002
  • Taklihwangki-Tang was a drug that treated carbuncle and cellulitis. So, the purpose of this Study was to investigate effect of Taklihwangki-Tang on the anti-cancer and proliferation of immunocytes, nitric oxide(NO) production of peritoneal macrophages. We used Taklihwangki-Tang extract(THT) with freeze-dried, 8wks-old male mice and cancer cell lines(L1210, S-180) for this Study. The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). The results of this Study were obtained as follow ; THT was showed cytotoxicity on the L1210 and S-180 cell lines, increased proliferation of thymocytes. And the combined effects of THT and vincristine were became cytotoxicity of cancer cell lines and increased significantly proliferation of thymocytes. THT accelerated proliferation of thymocytes in normal mice, and decreased significantly proliferation of L1210 cells and accelerated significantly NO production of peritoneal macrophages in L1210 cells transplanted mice. This results suggest that THT inhibit proliferation of cancer cells by becoming immunocytes activity(NO production, proliferation of T-cell).

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Effect of Substrates on the Microbial Communities in a Microbial Electrolysis Cell and Anaerobic Digestion Coupled System (기질에 따른 미생물 전해 전지-혐기성 소화의 미생물 군집 특성)

  • LEE, CHAE-YOUNG;HAN, SUN-KEE
    • Journal of Hydrogen and New Energy
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    • v.30 no.3
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    • pp.269-275
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    • 2019
  • This study was conducted to evaluate the microbial communities in coupled system of a microbial electrolysis cell and an anaerobic digestion. Glucose, butyric acid, propionic acid and acetic acid were used as substrates. The maximum methane production and methane production rate of propionic acid respectively were $327.9{\pm}6.7mL\;CH_4/g\;COD$ and $28.3{\pm}3.1mL\;CH_4/g\;COD{\cdot}d$, which were higher than others. Microbial communities' analyses indicated that acetoclastic methangens were predominant in all systems. But the proportion of hydrogenotrophic methanogens was higher in the system using propionic acid as a substrate when compared to others. In coupled system of a microbial electrolysis cell and anaerobic digestion, the methane production was higher as the distribution of hydrogen, which was generated by substrate degradation, and proportion of hydrogenotrophic methanogens was higher.

Antibody Production in Plant Cell Cultures

  • Lee, James M.
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.06a
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    • pp.67-78
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    • 1995
  • Monoclonal antibodies (MoAbs) are a highly diversified class of proteins with major research and commercial applications such as diagnostics and therapeutics. Currently, the dominant method for producing MoAbs is through the hybridoma technique. However, this technique is slow, tedious, labor intensive, and expensive. The production of MoAbs in cultured transgenic plant cells can offer some advantages over that in the over that in the mammalian systems. The media to cultivate plant cells are well defined and inexpensive. Contamination by bacteria or fungi is easily monitored in plant tissue cultures. Furthermore, these contaminants are usually not potent pathogens to human beings. In our interdisciplinary research efforts, heavy chain monoclonal antibody (HC MAb) was inserted into Ti plasmid vector and transferred into A. tumefaciens for the transformation in tobacco cells. It was found that 76% of the transformants produced HC MAb. The presence of HC MAb in the cell membrane fraction indicated that the signal peptide was functional and efficient. The change of the HC MAb concentration during a batch culture followed a similar trend as dry cell concentration, indicating that the production of HC MAb was growth related. The long-term repeated subcultures of 11 cell lines showed that there was no obvious trend of neither the decrease nor the increase of the productivity with the repeated subcultures.

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Examination of the xanthosine response on gene expression of mammary epithelial cells using RNA-seq technology

  • Choudhary, Shanti;Li, Wenli;Bickhart, Derek;Verma, Ramneek;Sethi, R.S.;Mukhopadhyay, C.S.;Choudhary, Ratan K.
    • Journal of Animal Science and Technology
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    • v.60 no.7
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    • pp.18.1-18.12
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    • 2018
  • Background: Xanthosine treatment has been previously reported to increase mammary stem cell population and milk production in cattle and goats. However, the underlying molecular mechanisms associated with the increase in stem cell population and milk production remain unclear. Methods: Primiparous Beetal goats were assigned to the study. Five days post-partum, one mammary gland of each goat was infused with xanthosine (TRT) twice daily ($2{\times}$) for 3 days consecutively, and the other gland served as a control (CON). Milk samples from the TRT and CON glands were collected on the 10th day after the last xanthosine infusion and the total RNA was isolated from milk fat globules (MEGs). Total RNA in MFGs was mainly derived from the milk epithelial cells (MECs) as evidenced by expression of milk synthesis genes. Significant differentially expressed genes (DEGs) were subjected to Gene Ontology (GO) terms using PANTHER and gene networks were generated using STRING db. Results: Preliminary analysis indicated that each individual goat responded to xanthosine treatment differently, with this trend being correlated with specific DEGs within the same animal's mammary gland. Several pathways are impacted by these DEGs, including cell communication, cell proliferation and anti-microbials. Conclusions: This study provides valuable insights into transcriptomic changes in milk producing epithelial cells in response to xanthosine treatment. Further characterization of DEGs identified in this study is likely to delineate the molecular mechanisms of increased milk production and stem or progenitor cell population by the xanthosine treatment.

Change of Insulin-like Growth Factor Gene Expression in Chinese Hamster Ovary Cells Cultured in Serum-free Media

  • Park, Hong-Woo;An, Sung-Kwan;Choe, Tae-Boo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.319-324
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    • 2006
  • Although the sera used in animal cell culture media provide the macromolecules, nutrients, hormones, and growth factors necessary to support cell growth, it could also be an obstacle to the production of recombinant proteins in animal cell culture systems used in many sectors of the biotechnology industry. For this reason, many research groups, including our laboratory, have been trying to develop serum-free media (SFM) or serum-supplemented media (SSM) for special or multi-purpose cell lines. The Chinese hamster ovary (CHO) cell, for example, is frequently used to produce proteins and is especially valuable in the large-scale production of pharmaceutically important proteins, yet information about its genome is lacking. Also, SFMs have only been evaluated by comparing growth patterns for cells grown in SFMs with those grown in SSM or by measuring the titer of the target protein obtained from cells grown in each type of medium. These are not reliable methods of obtaining the type of information needed to determine whether an SFM should be replaced with an SSM. We carried out a cDNA microarray analysis to evaluate MED-3, an SFM developed in our laboratory, as a CHO culture medium When CHO cells were cultured in MED-3 instead of an SSM, several genes associated with cell growth were down-regulated, although this change diminished over time. We found that the insulin-like growth factor (IGF) gene was representative of the proteins that were down-regulated in cells cultured in MED-3. When several key supplements - including insulin, transferrin, ethanolamine, and selenium - were removed from MED-3, the IGF expression was consistently down- regulated and cell growth decreased proportionately. Based on these results, we concluded that when an SFM is used as a culture medium, it is important to supplement it with substances that can help the cells maintain a high level of IGF expression. The data presented in this study, therefore, might provide useful information for the design and development of SFM or SSM, as well as for the design of genome-based studies of CHO cells to determine how they can be used optimally for protein production in pharmaceutical and biomedical research.

Effects of Baekgumhwan administration on immune-function in ICR mice stressed by electric footshock (백금환(白金丸)의 경구 투여가 전기자극 스트레스를 받은 mouse의 비장에 존재하는 면역 세포 분획과 사이토카인 생성에 미치는 영향)

  • Joo, Seung-Gyun;Kim, Geun-Woo;Koo, Byung-Soo;Shim, Sang-Min
    • Journal of Oriental Neuropsychiatry
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    • v.13 no.1
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    • pp.39-52
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    • 2002
  • The present experiments were designed to study the influence of Baekgumhwan on immune function of ICR mice under stress condition. Baekgumhwan was orally administered to the mice for 15days. on the 11th day the mice subjected to electric footshock for 5days(2 session a day, 11 footshocks a 31 min-session). B/T cell populations in splenocytes were studied by FACS analysis and cytokines($IFN-{\gamma}$ rand IL-10) production of the mouse splenocytes treated with PHA were studied by sandwich ELISA assay on the 15th day. The results were as follows. 1. After electric footshock, mice became sluggish and crowded to one side of the cage. Increased B/T cell populations in splenocytes were observed. These results confirm that electric footshock caused stress inducing immunological and behavioral changes in ICR mice. 2. Baekgumhwan administration without stress increase B cell populations in splenocytes, but T cell populations and cytokines($IFN-{\gamma}$ and IL-10) production of the mouse splenocytes treated with PHA maintain as similar levels as in the normal group. 3. Baekgumhwan administration with stress significantly antagonized the effect of electric footshock on behavior, increased B cell populations in splenocytes, so maintain as similar levels as in the normal group. cytokines($IFN-{\gamma}$ rand IL-10) production of the mouse splenocytes treated with PHA maintain as similar levels as in the normal group and T cell populations in splenocytes were increased as stress control.

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The Effects of Jeondo-san on Anti-Inflammation and Anti-Propionibacterium acnes (전도산(顚倒散)이 여드름 유발균과 염증에 미치는 영향)

  • Choi, Kwan-Ho;Seo, Hyeong-Sik
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.20 no.2 s.33
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    • pp.89-101
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    • 2007
  • Objectives : This study was carried out to investigate the effects of Jeondo-San(JDS) on anti-Inflammation and anti-Propionibacterium acnes. Methods : The effects of JDS on anti-Inflammation and anti-Propionibacterium acnes were measured by the cytotoxicity of Raw 264.7 cell, the inhibition for NO, $TNF-{\alpha}$, $PGE_2$, iNOS and COX-2, the blocking $NF-{\kappa}B$ into nucleus and the sterilizing power for Propionibacterium acnes. Results : 1. All concentrations of JDS has no cytotoxicity in Raw 264.7 cell. 2. All concentrations of JDS inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 3. All concentrations of JDS did not significantly inhibit the production of $TNF-{\alpha}$ in the Raw 264.7 cell stimulated with LPS. 4. All concentrations of JDS inhibited the production of $PGE_2$ in the Raw 264.7 cell stimulated with LPS. 5. All concentrations of JDS did not inhibit the expression of COX-2 but concentrations of 50\;{\mu}g/ml$, 100\;{\mu}g/ml$ JDS inhibited iNOS expression in the Raw 264.7 cell stimulated with LPS. 6. Concentrations of 50\;{\mu}g/ml$, 100\;{\mu}g/ml$ JDS has the effect of blocking $NF-{\kappa}B$ into nucleus in LPS-induced macrophage Raw 264.7 cell. 7. All concentrations of IDS did not have the inhibitory effect of Propionibactrium acnes. Conclusions : The present date suggest that JDS has a effect on the stage of inflammation of acne.

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Effect of Carbon dioxide in Fuel on the Performance of PEM Fuel Cell (연료중의 이산화탄소 불순물에 의한 연료전지 성능변화 연구)

  • Seo, Jung-Geun;Kwon, Jung-Taek;Kim, Jun-Bom
    • 한국신재생에너지학회:학술대회논문집
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    • 2007.11a
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    • pp.184-187
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    • 2007
  • Hydrogen could be produced from any substance containing hydrogen atoms, such as water, hydrocarbon (HC) fuels, acids or bases. Hydrocarbon fuels couold be converted to hydrogen-rich gas through reforming process for hydrogen production. Even though fuel cell have high efficiency with pure hydrogen from gas tank, it is more beneficial to generate hydrogen from city gas (mainly methane) in residential application such as domestic or office environments. Thus hydrogen is generated by reforming process using hydrocarbon. Unfortunately, the reforming process for hydrogen production is accompanied with unavoidable impurities. Impurities such as CO, $CO_2$, $H_2S$, $NH_3$, and $CH_4$ in hydrogen could cause negative effects on fuel cell performance. Those effects are kinetic losses due to poisoning of electrode catalysts, ohmic losses due to proton conductivity reduction including membrane and catalyst ionomer layers, and mass transport losses due to degrading catalyst layer structure and hydrophobic property. Hydrogen produced from reformer eventually contains around 73% of $H_2$, 20% or less of $CO_2$, 5.8% of less of $N_2$, or 2% less of $CH_4$, and 10ppm or less of CO. Most impurities are removed using pressure swing adsorption (PSA) process to get high purity hydrogen. However, high purity hydrogen production requires high operation cost of reforming process. The effect of carbon dioxide on fuel cell performance was investigated in this experiment. The performance of PEM fuel cell was investigated using current vs. potential experiment, long run (10 hr) test, and electrochemical impedance measurement when the concentrations of carbon dioxide were 10%, 20% and 30%. Also, the concentration of impurity supplied to the fuel cell was verified by gas chromatography (GC).

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Expression Vectors for Human-mouse Chimeric Antibodies

  • Xiong, Hua;Ran, Yuliang;Xing, Jinliang;Yang, Xiangmin;Li, Yu;Chen, Zhinan
    • BMB Reports
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    • v.38 no.4
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    • pp.414-419
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    • 2005
  • The production of recombinant antibodies has been generally recognized as time-consuming and labor-intensive. The aim of our study is to construct mammalian expression vectors containing the cDNA encoding the human constant regions and murine variable regions to massively and cost-effectively produce full-length chimeric antibodies. Unique restriction sites flanking the Ig variable region were designed to allow for the replacement of variable regions generated by PCR. Western blot analysis of the chimeric antibodies revealed that the expressed products were of the predicted size, structure and specificity. The usefulness of the vectors was confirmed by construction of human-mouse chimeric antibody-HCAb which secretes murine antibody against the human colorectal cancer. Selected in medium containing gradually increasing methotrexate (MTX), clones with increased expression of the product gene can be efficiently generated. The secretion of recombinant chimeric antibody-HCAb yielded $30\;pg\;cell^{-1}\;day^{-1}$ at $10^{-6}\;M$ MTX. With this high-level expression from pools, the convenient and rapid production of over 100 milligram amounts per liter of recombinant antibodies may be achieved, which indicates the significant roles of pYR-GCEVH and pYR-GCEVL in the production of chimeric antibodies.