• Title/Summary/Keyword: cell production

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Development of Good Manufacturing facility for Radiopharmaceuticals (우수방사성의약품 생산시설 개발)

  • Shin, Byung-Chul;Choung, Won-Myung;Park, San-Hyun;Lee, Kyu-Il;Park, Kyung-Bae;Park, Jin-Ho
    • Journal of Pharmaceutical Investigation
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    • v.33 no.2
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    • pp.145-149
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    • 2003
  • Manufacturing facilities of the pharmaceuticals must meet certain level of the cleanness required so that foreign substances such as dust, moisture, heat, microorganism, or virus do not contaminate the product. In case of radiopharmaceuticals for medical treatment and diagnosis, not only should the operators and environment be protected from radiation but also need to be isolated from the foreign contaminant. Therefore, manufacturing facilities for radiopharmaceuticals must satisfy the design standards of both hot cell and clean room which are specified by GMP. However, standards of maintaining negative pressure for preventing spread of radioactive contaminant in isolated facilities conflict with the standards of maintaining positive pressure for keeping cleanness. To solve this problem, air pressure of hot cell was designed lower than in the adjacent area to meet standards of the radiation safety. To keep higher cleanness in certain part of the hot cell for filling, minimal relative positive pressure allows. In order to effectively maintain the cleanness that is required for production of Tc-99m generator, which takes 70% of whole demand of radiopharmaceuticals, the rooms placed in each side of production room are used as a buffer area and three lead hot cells are installed in production room. In this research, we established the appropriate engineered design concept for Tc-99m generator manufacturing facility, which satisfies both GMP cleanness standard for preventing particles, bacteria, other contaminants and the regulations of radiation safety for supervising and controlling the amount of radiation exposure and exhausted radioactivity. And the concept of multi-barrier buffer zones is introduced to apply negative air pressure for hot cell with first priority and to continue relative positive air pressure for clean room.

Anti-proliferative Activity of T-bet

  • Oh, Yeon Ji;Shin, Ji Hyun;Won, Hee Yeon;Hwang, Eun Sook
    • IMMUNE NETWORK
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    • v.15 no.4
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    • pp.199-205
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    • 2015
  • T-bet is a critical transcription factor that regulates differentiation of Th1 cells from $CD4^+$ precursor cells. Since T-bet directly binds to the promoter of the IFN-${\gamma}$ gene and activates its transcription, T-bet deficiency impairs IFN-${\gamma}$ production in Th1 cells. Interestingly, T-bet-deficient Th cells also display substantially augmented the production of IL-2, a T cell growth factor. Exogenous expression of T-bet in T-bet deficient Th cells rescued the IFN-${\gamma}$ production and suppressed IL-2 expression. IFN-${\gamma}$ and IL-2 reciprocally regulate Th cell proliferation following TCR stimulation. Therefore, we examined the effect of T-bet on Th cell proliferation and found that T-bet deficiency significantly enhanced Th cell proliferation under non-skewing, Th1-skewing, and Th2-skewing conditions. By using IFN-${\gamma}$-null mice to eliminate the anti-proliferative effect of IFN-${\gamma}$, T-bet deficiency still enhanced Th cell proliferation under both Th1- and Th2-skewing conditions. Since the anti-proliferative activity of T-bet may be influenced by IL-2 suppression in Th cells, we examined whether T-bet modulates IL-2-independent cell proliferation in a non-T cell population. We demonstrated that T-bet expression induced by ecdysone treatment in human embryonic kidney (HEK) cells increased IFN-${\gamma}$ promoter activity in a dose dependent manner, and sustained T-bet expression considerably decreased cell proliferation in HEK cells. Although the molecular mechanisms underlying anti-proliferative activity of T-bet remain to be elucidated, T-bet may directly suppress cell proliferation in an IFN-${\gamma}$- or an IL-2-independent manner.

Production of Bovine Transgenic Embryos Derived from Non-transfected and Transfected Adult Cells (외부유전자가 도입된 체세포를 이용한 소 형질전환 복제란 생산)

  • J. K. Cho;M.M.U. Bhuiyan;G. Jang;Park, E. S.;J. M. Lim;S. K. Kang;Lee, B. C.;W. S. Hwang
    • Journal of Embryo Transfer
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    • v.17 no.2
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    • pp.109-115
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    • 2002
  • The present study was conducted for the production of transgenic cloned cows those secrete human lactoferricin into milk by somatic cell nuclear transfer (NT). To estimate detrimental effects of gene transfection on transgenic cloned embryo production, development rates of NT embryos were compared between transfected and non-transfected cumulus and ear fibroblast cells. An expression plasmid for human lactofericin (pbeta-LFC) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and human lactoferricin target gene into a pcDNA3 plasmid. Two bovine somatic cell lines (cumulus cell and ear fibroblast) were established and transfected with the expression plasmid using a liposomal transfection reagent, Fugene6 as a carrier. Cumulus cell and ear fibroblast were transfected at the passage of 2 to 4, trypsinized and GFP-expressing cells were randomly selected and used for somatic cell NT. Developmental competences (rates of fusion, cleavage, and blastocyst formation) in bovine transgenic somatic cell NT embryos reconstructed with non-transfectecd cells were significantly higher than those from transfected cells in cumulus cell and ear fibroblast (P<0.05). This study indicated that transfection of done. cell has detrimental effect on embryo development in bovine transgenic NT.

Effects of Fucoidan on Neuronal Cell Proliferation: Association with NO Production through the iNOS Pathway

  • Lee, Hye-Rim;Do, Hang;Lee, Sung-Ryul;Sohn, Eun-Soo;Pyo, Suhk-Neung;Son, Eun-Wha
    • Preventive Nutrition and Food Science
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    • v.12 no.2
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    • pp.74-78
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    • 2007
  • Fucoidan, that is high-molecular-weight sulfated polysaccharides extracted from brown seaweeds has been shown to elicit various biological activities. Here, we investigated the effects of fucoidan on cell proliferation and nitric oxide (NO) production in neuronal blastoma cell (SH-SY5Y). In the present study, we demonstrated that fucoidan treatment resulted in increase of cell proliferation and NO production. When cells were treated with amyloid-${\beta}$ (A${\beta}$) in the absence or presence of fucoidan, fucoidan recovered the cell viability decreased by A${\beta}$ peptides. To further determine whether nitric oxide synthase (NOS) is involved in proliferative effect of fucoidan, cells were treated with NOS inhibitors in the absence or presence of fucoidan. Selective constitutive nitric oxide synthase (cNOS) inhibitor, diphenylene iodonium chloride (DPI), caused a decrease of cell viability, whereas cell viability was increased by specific inducible nitric oxide synthase (iNOS) inhibitor, S-methylisothiourea (SMT), in the fucoidan-untreated cells. Treatment with fucoidan inhibited the cell viability decreased in DPI-exposed cells. In contrast, fucoidan had no effect on cell growth in SMT-treated cells, indicating that cNOS may not play a role in the proliferation of fucoidan-treated cells. The present data suggest that fucoidan has proliferative and neuroprotective effects and these effects may be associated with iNOS.

ELECTRO-MICROSCOPE BASED 3D PLANT CELL IMAGE PROCESSING METHOD

  • Lee, Choong-Ho;Umeda Mikio;Takesi Sugimoto
    • Proceedings of the Korean Society for Agricultural Machinery Conference
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    • 2000.11b
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    • pp.227-235
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    • 2000
  • Agricultural products are easily deformable its shape because of some external forces. However, these force behavior is difficult to measure quantitatively. Until now, many researches on the mechanical property was performed with various methods such as material testing, chemical analysis and non-destructive methods. In order to investigate force behavior on the cellular unit of agricultural products, electro-microscope based 3D image processing method will contribute to analysis of plant cells behavior. Before image measurement of plant cells, plant sample was cut off cross-sectioned area in a size of almost 300-400 ${\mu}$ m units using the micron thickness device, and some of preprocessing procedure was performed with fixing and dyeing. However, the wall structure of plant cell is closely neighbor each other, it is necessary to separate its boundary pixel. Therefore, image merging and shrinking algorithm was adopted to avoid disconnection. After then, boundary pixel was traced through thinning algorithm. Each image from the electro-microscope has a information of x,y position and its height along the z axis cross sectioned image plane. 3D image was constructed using the continuous image combination. Major feature was acquired from a fault image and measured area, thickness of cell wall, shape and unit cell volume. The shape of plant cell was consist of multiple facet shape. Through this measured information, it is possible to construct for structure shape of unit plant cell. This micro unit image processing techniques will contribute to the filed of agricultural mechanical property and will use to construct unit cell model of each agricultural products and information of boundary will use for finite element analysis on unit cell image.

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Vinegar Production by Acetobacter aceti Cell Immobilized in Calcium Alginate (Calcium Alginate로 고정화된 Acetobacter aceti에 의한 식초생산)

  • 유익제;박기문유연우최춘언
    • KSBB Journal
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    • v.5 no.2
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    • pp.167-173
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    • 1990
  • This study is to investigate for obtaining the operating conditions of continuous vinegar production using fluidized bed reactor by Acetobacter aceti cell immobilized in Ca-alginate gel. The optimum conditions obtaining by batch fermentation using fluidized bed reactor were as follows; The fermentation temperature and aeration rate were 3$0^{\circ}C$ and 1.0VVM and the initial concentration of ethanol and acetic acid in medium were 33g/l and 27g/l respectively. The amount of bead used was 25%(w/v). The overall acetic acid productivities of batch fermentations by free cell and immobilized cell were 0.31g/l-hr and 0.48g/l-hr, respectively, at the final acetic acid concentration of 50g/l. In the continuous vinegar production using fluidized bed reactor by immobilized cell under optimum conditions, it was possible to produce 23g/l acetic acid continuously up to 90 days with maximum acetic acid productivity of 2.76g/l-hr at dilution rate 0.12hr-1.

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Effects of nitrogen and organic carbon sources on growth and lipid production of Chlorella sp. KR-1 in flask cultures (플라스크 배양에서 Chlorella sp. KR-1의 균체 성장 및 지질 생산에 대한 질소원 및 유기탄소원의 영향)

  • Lee, Ja-Youn;Seo, Kyoung Ae;Oh, You-Kwan
    • Journal of Marine Bioscience and Biotechnology
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    • v.6 no.2
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    • pp.110-117
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    • 2014
  • Recently microalgae have been proposed as a promising biodiesel feedstock, owing to their higher lipid productivity and non-arable land based cultivation system. Biomass and lipid productivities of microalgae are largely affected by various environmental and nutritional factors. In this study, the effects of nitrogen (nitrate and ammonium) and organic carbon (glucose and glycerol) sources on the cell growth and lipid production of Chlorella sp. KR-1 were examined in flask cultures. Under autotrophic culture conditions for 15 days, overall cell growth and lipid (fatty acid methyl ester, FAME) production with nitrate were better than those of ammonium, resulting in 1.06 g cell/L and 333 mg FAME/L, respectively. Maximal intracellular lipid contents (348 - 352 mg FAME/g cell) were observed at low concentrations of 1 mM for both nitrate and ammonium. In the supply of light, addition of glucose in the range of 1 - 20 g/L showed higher cell densities than the autotrophic cell growth condition. Higher lipid accumulation of 375 mg FAME/g cell could achieved at 5 g glucose/L albeit of relatively short incubation of 7 days. With glycerol, intracellular lipid contents were ~1.9 times lower than glucose cases although similar cell growths were observed for both carbon sources.

Combined Effect of Catholyte Gap and Cell Voltage on Syngas Ratio in Continuous CO2/H2O Co-electrolysis

  • Ha, Min Gwan;Na, Youngseung;Park, Hee Young;Kim, Hyoung-Juhn;Song, Juhun;Yoo, Sung Jong;Kim, Yong-Tae;Park, Hyun S.;Jang, Jong Hyun
    • Journal of Electrochemical Science and Technology
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    • v.12 no.4
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    • pp.406-414
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    • 2021
  • Electrochemical devices are constructed for continuous syngas (CO + H2) production with controlled selectivity between CO2 and proton reduction reactions. The ratio of CO to H2, or the faradaic efficiency toward CO generation, was mechanically manipulated by adjusting the space volume between the cathode and the polymer gas separator in the device. In particular, the area added between the cathode and the ion-conducting polymer using 0.5 M KHCO3 catholyte regulated the solution acidity and proton reduction kinetics in the flow cell. The faradaic efficiency of CO production was controlled as a function of the distance between the polymer separator and cathode in addition to that manipulated by the electrode potential. Further, the electrochemical CO2 reduction device using Au NPs presented a stable operation for more than 23 h at different H2:CO production levels, demonstrating the functional stability of the flow cell utilizing the mechanical variable as an important operational factor.

Evaluation of Macroporous and Microporous Carriers for CHO-K1 Cell Growth and Monoclonal Antibody Production

  • Rodrigues, Maria Elisa;Costa, Ana Rita;Fernandes, Pedro;Henriques, Mariana;Cunnah, Philip;Melton, David W.;Azeredo, Joana;Oliveira, Rosario
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1308-1321
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    • 2013
  • The emergence of microcarrier technology has brought a renewed interest in anchorage-dependent cell culture for high-yield processes. Well-known in vaccine production, microcarrier culture also has potential for application in other fields. In this work, two types of microcarriers were evaluated for small-scale monoclonal antibody (mAb) production by CHO-K1 cells. Cultures (5 ml) of microporous Cytodex 3 and macroporous CultiSpher-S carriers were performed in vented conical tubes and subsequently scaled-up (20 ml) to shake-flasks, testing combinations of different culture conditions (cell concentration, microcarrier concentration, rocking methodology, rocking speed, and initial culture volume). Culture performance was evaluated by considering the mAb production and cell growth at the phases of initial adhesion and proliferation. The best culture performances were obtained with Cytodex 3, regarding cell proliferation (average $1.85{\pm}0.11{\times}10^6$ cells/ml against $0.60{\pm}0.08{\times}10^6$ cells/ml for CultiSpher-S), mAb production ($2.04{\pm}0.41{\mu}g/ml$ against $0.99{\pm}0.35{\mu}g/ml$ for CultiSpher-S), and culture longevity (30 days against 10-15 days for CultiSpher-S), probably due to the collagen-coated dextran matrix that potentiates adhesion and prevents detachment. The culture conditions of greater influence were rocking mechanism (Cytodex 3, pulse followed by continuous) and initial cell concentration (CultiSpher-S, $4{\times}10^5$ cells/ml). Microcarriers proved to be a viable and favorable alternative to standard adherent and suspended cultures for mAb production by CHO-K1 cells, with simple operation, easy scale-up, and significantly higher levels of mAb production. However, variations of microcarrier culture performance in different vessels reiterate the need for optimization at each step of the scale-up process.

Immunomodulatory activities of crude polysaccharide fraction separated from Perilla frutescens Britton var. acuta Kudo (자소엽(Perilla frutescens Britton var. acuta Kudo) 조다당류 추출물의 면역활성 효과)

  • Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
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    • v.49 no.5
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    • pp.559-566
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    • 2017
  • This aim of this study was to examine the immunomodulatory activities of crude polysaccharides from Perilla frutescens Britton var. acuta Kudo (PCP) in mouse bone marrow-derived dendritic cells (BMDC) and splenocytes. The immunomodulatory activity was determined by cell viability, nitric oxide (NO) production, cell surface marker expression (CD 80/86 and MHC class I/II), and cytokine production in BMDC, and cell viability, and cytokine production in splenocytes. Cell proliferation and cytokine production (tumor necrosis factor; TNF-${\alpha}$, interleukin (IL)-6, IL-$1{\beta}$, and IL-12) tested in BMDC were significantly increased by PCP treatment. Additionally, the cell surface markers (CD 80/86, MHC class I/II) were highly increased by PCP treatment. For cytokine production in splenocytes, PCP treatment significantly increased the production of Th 1 cytokines [IL-2 and interferon (IFN)-${\gamma}$], but not Th 2 cytokines (IL-4). Therefore, PCP can induce immune cell activation and is a potential candidate for the development of nutraceuticals to boost the immune system.