• Title/Summary/Keyword: cell culture

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Microcarrier Culture of an Anchorage-dependent Cell Using Cytodex-3 (Cytodex-3를 이용한 부착성 동물세포의 미립담체 배양)

  • 김정회;최준호;웨이슈후
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.231-235
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    • 1989
  • Possibility of using microcarriers for the growth of a transformed human embryonic kidney cell line 293 was investigated. The cell grew well in a static culture such as T-flasks with medium of DME/F12 (3:1) mixture supplemented with 5% FBS, but it was most difficult to make the cells grow on microcarriers mainly due to the low attachment efficiency and poor spreading at initial stage of the culture. Consequently, 30-50% of the cells were lost upon inoculation into microcarrier suspension and significant fraction of the mirrocarrier became bald. The medium supplemented with the concentrated conditioned medium by hepatoma cell line HpG2 supported the active growth of the cells on microcarrier and the cells showed a very healthy and well spreading morphology. It was probable that some spreading and attachment factors of HpG2 conditioned medium were effective for 293 cells.

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Production of Cell Mass and Monacolin K from Monascus sp. on Rice Solid Culture (Monascus 속 균주의 균체 생산 및 고체배양에 의한 Monacolin K 생산)

  • 정혁준;유대식
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.160-166
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    • 2004
  • The optimal conditions for production of Monascus sp. KM100l cell mass on submerged culture and production of monacolin K on rice solid culture were investigated. An overproducing mutant of Monascus pigments, KM 1001 mutant, from Monascus purpureus KCCM60016 was selected by NTG treatment. The optimal medium for the production of KM100l mutant cell mass is instructed to be composed of 3% glucose, 2% yeast extract, 0.1 % KH$_2$PO$_4$, 0.05% The optimal conditions for production of Monascus sp. KM100l cell mass on submerged culture and production of monacolin K on rice solid culture were investigated. An overproducing mutant of Monascus pigments, KM 1001 mutant, from Monascus purpureus KCCM60016 was selected by NTG treatment. The optimal medium for the production of KM100l mutant cell mass is instructed to be composed of 3% glucose, 2% yeast extract, 0.1 % KH$_2$The optimal conditions for production of Monascus sp. KM100l cell mass on submerged culture and production of monacolin K on rice solid culture were investigated. An overproducing mutant of Monascus pigments, KM 1001 mutant, from Monascus purpureus KCCM60016 was selected by NTG treatment. The optimal medium for the production of KM100l mutant cell mass is instructed to be composed of 3% glucose, 2% yeast extract, 0.1 % $(KH_2PO_4$, 0.05% $MgSO_4{\cdot}7H_2O$, 0.2% L-asparagine, pH 4.5, and the optimal inoculum size and shaking speed were $1.5{\times}10^6$ spores/50 m1 medium and 150 rpm, respectively. On optimal conditions, 4.1 g/l of the cell mass was obtained at 28$^{\circ}C$ for 3 days. The mycelium were inoculated on 500 g of steamed rice using vinyl bag ($30.6{\times}44$ cm) and incubated at $30^{\circ}C$, 85% humidity for 21 days. Lactone form monacolin K was rapidly increased for 2 days and reached highest concentration of monacolin K (2,930 mg/kg) for 15 days, and monacolin K was decreased after 15 days.

Mathematical Analysis of a High Density Animal Cell Culture with a Spin-Filter (회전식 여과기를 이용한 고농도 동물세포배양의 수학적 해석)

  • 박흥우
    • KSBB Journal
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    • v.9 no.2
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    • pp.230-237
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    • 1994
  • Spin-filters are used as cell separation devices for achieving high cell density and high productivity in animal cell culture. We have proposed a model for the cell growth in a spin-filter perfusion culture and examined the effects on cell growth by several parameters including ammonia inhibition, specific growth rate, specific feeding rate, and cell retention. Results from computer simulation and sensitivity analysis indicate that the cell retention affects the cell growth mostly while there is a significant inhibition on cell growth by the ammonia accumulated during the culture. The specific feeding rate has minimal effects on cell growth, which is consistant with the fact that the cell growth with a step feeding is quite similar to that with a continuous feeding.

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Decusinol angelate inhibits UVB-induced MMP-l induction via Mitogen-activated Protein Kinase Pathway in human skin fibroblasts

  • Wang, Hye-Sung;Kang, Sun-Myung;Cho, Hong-Yon;Ho, Jin-Nyung;Kim, Ik-Hwan
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.337.3-338
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    • 2002
  • UV-B irradiation increases the synthesis of matrixmetalloproteinase-1 (MMP-1) that degrades skin collagen in human skin. In this work, we investigated the photoprotective effect of decursinol angelate (DEA) extracted Irom Angelica gigas on human skin libroblasts. DEA inhibited UVB-induced MMP-1 induction, which was conlirmed by western blot and ELISA. We examined upstream signal transduction pathway and the action mechanism of DEA on UVB induction of MMp in human skin fibroblasts. (omitted)

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A Spatial-domain Fourier Transform Infrared Spectrometer: Application for Analyte Measurement in Cell Culture Media

  • Jung, Byung-Jo
    • Journal of the Optical Society of Korea
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    • v.9 no.4
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    • pp.151-156
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    • 2005
  • A spatial-domain Fourier Transform (FT) infrared (IR) spectrometer coupled with a PtSi Schottky­barrier IR detector plane was developed in the spectral range of $2.0-2.5{\mu}m$ for noninvasive measurement of analyte concentrations in cell culture media during cell culture processing. A key optical component of the spectrometer is a Savart plate which is a birefringent polarizer generating coherent two rays for interfering. The spectral resolution of the spectrometer was determined as $71cm^{-1}$ (${\~}0.05{\mu}m$ at $2.5{\mu}m$). Clear IR fringe patterns were imaged on the IR detector plane. The feasibility of the spectrometer for our application was investigated by measuring absorbance spectra of glucose and fetal bovine serum (FBS) which are important compounds in cell culture media. Experiment results show that the spectral quality of glucose and FBS was comparable with the standard spectra acquired with a commercial FT-IR spectrometer, presenting the feasibility of the spectrometer to perform analyte measurement in cell culture media.

Development of the Three-Dimensional Perfusion Culture Technology for the Salivary Ductal Cells (타액선 도관세포의 관류 배양 기술 개발)

  • Kim, Ji Won;Kim, Jeong Mi;Choi, Jeong-Seok
    • International journal of thyroidology
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    • v.11 no.2
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    • pp.160-166
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    • 2018
  • Background and objectives: Salivary hypofunction is one of the common side effects after radioiodine therapy, and its pathophysiology is salivary ductal stenosis resulting from ductal cell injury. This study aimed to develop the functional culture environment of human parotid gland ductal cells in in vitro three-dimensional perfusion culture system. Materials and Methods: We compared plastic dish culture method and three-dimensional culture system containing Matrigel and nanofiber. Morphogenesis of reconstituted salivary structures was assessed by histomorphometry. Functional characteristics were assessed by immunohistochemistry and reverse transcription polymerase chain reaction (aquaporin 5, CK7, CK18, connexin 43, and p21). In addition, we designed the media perfusion culture system and identified higher rate of cell proliferation and expression of connexin 43 in perfusion system comparing to dish. Results: Human parotid ductal cells were well proliferated with the ductal cell characters under environment with Matrigel. In the presence of Matrigel, aquaporin 5, CK18 and connexin 43 were more expressed than 2D dish and 3D nanofiber setting. In the media perfusion culture system, ductal cells in 3D culture media showed higher cells count and connexin 43 expression compared to 2D dish. Conclusion: This in vitro ductal cell perfusion culture system using Matrigel could be used to study for radioiodine induced sialadenitis model in vivo.

Design and Performance of an Automated Bioreactor for Cell Culture Experiments in a Microgravity Environment

  • Kim, Youn-Kyu;Park, Seul-Hyun;Lee, Joo-Hee;Choi, Gi-Hyuk
    • Journal of Astronomy and Space Sciences
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    • v.32 no.1
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    • pp.81-89
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    • 2015
  • In this paper, we describe the development of a bioreactor for a cell-culture experiment on the International Space Station (ISS). The bioreactor is an experimental device for culturing mouse muscle cells in a microgravity environment. The purpose of the experiment was to assess the impact of microgravity on the muscles to address the possibility of long-term human residence in space. After investigation of previously developed bioreactors, and analysis of the requirements for microgravity cell culture experiments, a bioreactor design is herein proposed that is able to automatically culture 32 samples simultaneously. This reactor design is capable of automatic control of temperature, humidity, and culture-medium injection rate; and satisfies the interface requirements of the ISS. Since bioreactors are vulnerable to cell contamination, the medium-circulation modules were designed to be a completely replaceable, in order to reuse the bioreactor after each experiment. The bioreactor control system is designed to circulate culture media to 32 culture chambers at a maximum speed of 1 ml/min, to maintain the temperature of the reactor at $36{\pm}1^{\circ}C$, and to keep the relative humidity of the reactor above 70%. Because bubbles in the culture media negatively affect cell culture, a de-bubbler unit was provided to eliminate such bubbles. A working model of the reactor was built according to the new design, to verify its performance, and was used to perform a cell culture experiment that confirmed the feasibility of this device.

Secretory Production of hGM-CSF with a High Specific Biological Activity by Transgenic Plant Cell Suspension Culture

  • Kwon, Tae-Ho;Shin, Young-Mi;Kim, Young-Sook;Jang, Yong-Suk;Yang, Moon-Sik
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.135-141
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    • 2003
  • The human granulocyte-macrophage colony stimulating factor (hGM-CSF) gene was introduced into tobacco plants. The cell suspension culture was established from leaf-derived calli of the transgenic tobacco plants in order to express and secrete a biologically active hGM -CSF. The recombinant hGM-CSF from the transgenic plant cell culture (prhGM-CSF) was identified as a yield of about 180 ${\mu}$g/L in the culture filtrate, as determined by ELISA. The addition of 0.5 g/L polyvinylpyrrolidone (PVP) to the plant cell culture medium both stabilized the secreted prhGM-CSF and increased the level of production approximately 1.5-fold to 270 ${\mu}$g/L. The biological activity of the prhGM-CSF was confirmed by measuring the proliferation of the hGM-CSF-dependent cell line, TF-1. Interestingly, the specific activity of the prhGM-CSF was estimated to be approximately 2.7 times higher than that of a commercially available preparation from E. coli.

Effects of Sucrose level and Nitrogen Source on Fresh Weight and Anthocyanin Production in Cell Suspension Culture of 'Sheridan' Grape (Vitis spp.)

  • Kim, Seung-Heui;Kim, Seon-Kyu
    • Journal of Plant Biotechnology
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    • v.4 no.1
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    • pp.23-27
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    • 2002
  • To establish an in vitro mass production system of grape anthocyanin pigments through callus and cell suspension culture, the effects of nitrogen source and sucrose on fresh weight and anthocyanin production in cell suspension culture of 'Sheridan' grape level were studied. When the medium was devoid of $NO_3^-$, cell fresh weight was either remained stable (1% sucrose) or slightly decreased with culture time (2,3, and 4% sucrose). When $NH_4^-$ was lacking, 3% sucrose was most favorable for cell growth. When $NH_4^-$ was supplied as N source, the anthocyanin content of 2% sucrose containing medium was maintained 2 times higher than other levels till day 8 in culture, then that of 3 and 4% sucrose which peaked at day 12 thereafter. The anthocyanin content was low than $NO_3^-$-free media. Total anthocyanin content in $NH_4^-$-free medium was just about a half of that of $NH_4^+$ medium. Anthocyanin production of 2% sucrose in $NH_4^+$ medium was maintained about 3-fold till day 8, then decreased thereafter. In $NH_4^+$ medium, pH decreased gradually with final pH of 3.5 to 4.0, while pH in $NH_4^+$-free medium increased with final pH of 6.5 to 7.5.

Detection of Human Adenoviruses and Enteroviruses in Korean Oysters Using Cell Culture, Integrated Cell Culture-PCR, and Direct PCR

  • Choo Yoe-Jin;Kim Sang-Jong
    • Journal of Microbiology
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    • v.44 no.2
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    • pp.162-170
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    • 2006
  • Oysters are known to be carriers of food-born diseases, but research on viruses in Korean oysters is scarce despite its importance for public health. We therefore tested oysters cultivated in Goheung, Seosan, Chungmu, and Tongyeong, for viral contamination using cell culture and integrated cell culture PCR (ICC-PCR) with Buffalo green monkey kidney (BGMK) and human lung epithelial (A549) cells. Additional screens via PCR, amplifying viral nucleic acids extracted from oysters supplemented our analysis. Our methods found 23.6 %, 50.9 %, and 89.1 % of all oysters to be positive for adenoviruses when cell culture, ICC-PCR, and direct PCR, respectively, was used to conduct the screen. The same methodology identified enteroviruses in 5.45%, 30.9%, and 10.9% of all cases. Most of the detected enteroviruses (81.3%) were similar to poliovirus type 1; the remainder resembled coxsackievirus type A1. A homology search with the adenoviral sequences revealed similarities to adenovirus subgenera C (type 2, 5, and 6), D (type 44), and F (enteric type 40 and 41). Adenovirus-positive samples were more abundant in A549 cells (47.3%) than in BGMK cells (18.2 %), while the reverse was true for enteroviruses (21.8 % vs. 14.5 %). Our data demonstrate that Korean oysters are heavily contaminated with enteric viruses, which is readily detectable via ICC-PCR using a combination of A549 and BGMK cells.