• Title/Summary/Keyword: 연속세포분리

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Effects of Trypsin, Collagenase and Dimethyl Sulfoxide on Dissociation of Rat Heart Cells (배양을 위한 심근세포분리에 미치는 Trypsin, Collagenase와 Dimethyl Sulfoxide의 영향)

  • Park, Chang-Woo;Lee, Yung-Chang
    • Journal of Yeungnam Medical Science
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    • v.4 no.1
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    • pp.17-23
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    • 1987
  • New born rat heart cells were dissociated using trypsin and/or collegenase to elucidate the dissociation efficiency of these two enzymes. And the effect of dimethyl sulfoxide during and immediately after cell dissociation was also investigated to clarify the so-called protective activity of dimethyl sulfoxide on cell performance. The results can be summarized as follows. 1. Cold trypsin 18 hours pretreatment followed by warm collagenase treatment resulted best cell viability and cell yield. 2. Single, warm trypsin treatment gave the poorest result. 3. Dimethyl sulfoxide did not seem to play any protective role during or immediately after rat heart cell dissociation. It had very damaging effect on rat heart cells.

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A Continuous Cell Separator Based on Gravity and Buoyant Forces in Fluids of Dissimilar Density (서로 다른 밀도의 유체 내 바이오 물질이 받는 중력과 부력 차를 이용한 연속적 세포 분리기)

  • Oh, Ae-Gyoung;Lee, Dong-Woo;Cho, Young-Ho
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.36 no.4
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    • pp.391-395
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    • 2012
  • We present a continuous cell separator that achieves density-dependent and size-independent cell separation based on the net force of gravity and buoyancy forces on the cells in dissimilar density fluids. Previous cell separators are, based on the size or dielectrophoretic property of the cells and, are suitable for size-dependent and density-independent cell separation. However, these properties can make it difficult to collect the same types of cells with the same density but with size variations. The present separator, however, is capable of collecting the same types of cells based on the cell density in the fluid. Regardless of cell size, the proposed chip isolates low density cells, (white blood cells, or WBCs) at the upper outlet while obtaining high-density cells (red blood cells, or RBCs) from the lower outlet based on density. Efficiency levels for separation of WBCs and RBCs were $90.9{\pm}9.1%$ and $86.4{\pm}1.99%$, respectively. The present separator therefore has the potential for use in the pretreatment of whole blood.

Development of Continuous Clean Bioprocess for Kasugamycin Production (Kasugamycin 생산을 위한 연속 청정생물공정 개발)

  • Kim, Chang Joon;Park, Sun Ok;Chang, Yong Keun;Chun, Gie-Taek;Lee, Jong-Dae;Kim, Sangyong
    • Clean Technology
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    • v.4 no.1
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    • pp.45-59
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    • 1998
  • Continuous immobilized-cell culture was carried out for the production of kasugamycin, a secondary metabolite by a filamentous bacteria, Streptomyces kasugaensis, with an intention of reducing waste generation. A sporulation medium was developed for production of bulk amounts of spores, and the spores were entrapped into celite biosupports for immobilization. It was possible to effectively keep the immobilized-cells inside the reactor during the continuous culture by an efficient immobilized cell separator of decantor type on the outlet of the fermentor. Using this continuous immobilized-cell fermentor system, we investigated the effects of feed substrate and phosphate concentrations on kasugamycin production and chemical oxygen demand(COD). Comparing with the conventional suspended-cell batch culture, the kasugamycin productivity was observed to increase by 2.5 times, whereas COD per unit kasugamycin production decreased by 2.3 times in the continuous immobilized-cell culture. Based on these results, the continuous immobilized-cell system was considered to be a cleaner bioprocess than the conventional batch suspended-cell system.

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전분당 부산물로부터 유기산 생산 및 농축에 관한 연구 : II. 유기산의 대량생산을 위한 연속발효 및 액-액 추출

  • Kim, Dae-Sik;Choe, Cheol-Ho;Lee, Ui-Sang
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.121-124
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    • 2000
  • Organic acids were produced from wastes streams in food industries by cell-recycle fermentation using Propionibacterium acidipropionici ATCC 4965. As a results of continuous fermentation, maximum productivity was 3.32g organic acid/L/hr at the dilution rate of 0.2/hr. Compared to batch fermentation, maximum productivity was improved by as much as 13 times and cell mass production was increased by as much as 22 times. The diluted organic acids in the fermenter were selectively separated by liquid-liquid extraction using 30%(w/w) trioctylamine(TOA) dissolved in methylisobutylketone(MIBK). The degree of extraction was reached above 90% for both acetic and propionic acid through repeated extraction of organic acids in fermentation broth.

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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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역미셀을 이용한 참치내장으로부터 단백질 추출

  • 윤성옥;전병수
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.98-99
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    • 2000
  • 최근 유전자 공학의 급속한 발전으로 인하여 유전자 조작을 통한 생화합물의 대량 생산, 세포배양에 의한 생화학제품(아미노산, 단백질, 핵산)의 생산이 크게 진보하고 있으나, 이에 비해 생화학제품을 효율적으로 분리 및 정제하는 기술은 상대적으로 발전하지 못했다. 생화합물의 분리 방법은 전통적으로 유기용매에 의한 침전법, 염석법, 탈염법, 크로마토그래피법, 전기영동법, 흡착제를 이용한 흡착법등을 사용하는데 이 방법들은 소규모 및 회분식 공정으로 이루어져 대형화에 따른 분리 공정의 효율이 감소하는 어려움이 있었다. 따라서 연속공정으로서 대형화가 쉽고, 효율적인 생물분리공정의 개발이 요구되고 있다. (중략)

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A Continuous Cell Separation Chip Using Hydrodynamic Dielectrophoresis Process (유체동역학적 유전영동법을 이용한 극소형 연속 세포분리기)

  • Doh Il;Cho Young-Ho
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.29 no.1 s.232
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    • pp.53-58
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    • 2005
  • We present a high-throughput continuous cell separation chip using hydrodynamic dielectrophoresis (DEP) process. The continuous cell separation chip uses three planar electrodes in a separation channel, where the positive DEP cells are moved away from the central streamline while the negative DEP cells remain in the central streamline. In the experimental study, we use the mixture of viable (live) and nonviable (dead) yeast cells in order to obtain the continuous cell separation conditions. For the conditions of the electric fields frequency of 5MHz and the medium conductivity of $5{\mu}S/cm$, the fabricated chip performs a continuous separation of the yeast cell mixture at the varying flow-rate in the range of $0.1{\sim}{\mu{\ell}/min$.; thereby, resulting in the purity ranges of $95.9{\sim}97.3\%\;and\;64.5{\sim}74.3\%$ respectively for the viable and nonviable yeast cells. present chip demonstrates the constant cell separation performance for varying mixture flow-rates.

Isolation and cultivation of follicle constituting cells from human hair follicles

  • Shin, Youn-Ho;Seo, Young-Kwon;Lee, Doo-Hoon;Yoo, Bo-Young;Song, Kye-Yong;Seo, Seong-Jun;Whang, Sung-Joo;Kim, Young-Jin;Yang, Eun-Kyung;Park, Chang-Seo;Chang, Ih-Seop;Park, Jung-Keug
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.365-368
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    • 2003
  • Hair follicles develop as a result of epithelial-mesenchymal interactions between epidermal keratinocytes and dermal cells. Moreover hair follicles constitute multiple cells that influence hair follicle development and cyclic activity. We isolated some cells using explantation and enzymatic digestion method from human scalp hair follicles. So we could culture some follicular cells, such as outer root sheath (ORS) cells, dermal papilla (DP) cells, dermal sheath (DS) cells, matrix cells and melanocyte.

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Analysis of the Proteins Accumulated During Cold Treatment in Intermolecular Space of Barley (저온에서 세포밖 공간에 축적되는 보리 단백질)

  • 황철호
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.1
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    • pp.15-28
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    • 1995
  • In order to identify an antifreeze proteins responsible for freeze-tolerance in barley the proteins accumulated in extracellular space during cold acclimination were extracted and analyzed. After 42 days cold treatment there were several proteins sized of 70, 21, 16, 14 KDa increased in their amount accumulated in extracellular space. In addition, continuously sized polypeptides smaller than 10 KDa were found to be increased in their amount as cold treatment prolongs. Since these proteins were not detectable in total leaf protein extract it appears that the procedure used to isolate extracellular extract was valid. A similarity in profile of the extracellular proteins isolated from barley and rye may indicate a possibility for these proteins to be an antifreeze proteins since the same extract from rye was reported to show an antifreezing activity.

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Purification of Nitric Oxide Synthase from Bovine Pancreas

  • Nam, Suk-Woo;Seo, Dong-Wan;Lee, Young-Jin;Sung, Dae-Seok;Han, Jung-Whan;Lee, Hyang-Woo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.184-184
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    • 1996
  • Nitric Oxide Synthase(NO synthase: EC.1.14.13.39)는 생체내에서 L-arginine을 기질로 하여 nitric oxide(NO)와 L-citrulline의 생성을 매개하는 효소로서 뇌, 간장, 신장, 체장등 대부분의 주요장기와 근육세포, 신경세포 등 거의 모든 조직에 분포하고 있다. NO synthase에 의해 생성되는 NO는 혈관이완작용, 신경전달 물질로서의 작용, 면역 담당세포에서의 세포 독작용 등 많은 생리현상에 중요한 역할을 하는 것으로 알려져 있다. 특히 체장에서는 췌외분비 기능의 항진에 있어 세포내 cGMP level의 변동이 NO와 연관된다는 사실에 주목하고 있으며 본연구실에서도 이에 관한 연구가 진행중이다. 따라서 본 연구에서는 소 췌조직의 100,000$\times$g cytosol을 효소원으로 하여 다음과 같이 NO synthase의 분리, 정제를 시행하였다. Ammonium sulfate로 30%(176g solid ammonium sulfate/$\ell$) 포화, 침전 후 2',5'-ADP agarose 및 calmodulin-agarose affinity chromatography를 연속적으로 시행하여 NO synthase를 분리하였으며 electrophoresis상에서 약 160kd의 분자량을 나타내었다.

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