• Title/Summary/Keyword: 무혈청배지

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Pulse-Feeding of Serum Free Media for Enhancing Monoclonal Antibody Production under Perfusion Operation (연속배양에서 단일항체 생산성 향상을 위한 무혈청 배지의 단계적 유입)

  • 강재구;박형환;이현용
    • Microbiology and Biotechnology Letters
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    • v.18 no.1
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    • pp.61-65
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    • 1990
  • Lectin related inducer can enhance IgG$_1$ production rate from murine hybridoma cells by employing step-feeding of serum free media with producing about 40 mg/$\ell$ of monoclonal antibodies. This step-feeding perfusion process also proves to be able to cutivate animal cells when serum free media can not support the growth of these cells in perfusion process, as well as to improve production rate. This process yields about 28 x 10$^{-10}$ mg of MAb/cells/h compared to 11.1 x 10$^{-10}$ and 4.0 x 10$^{-11}$ mg/cells/h for perfusion process and batch cultivation with 10% serum containing media, respectively.

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Characteristics of Recombinant CHO Cell Growth and Erythropoietin Production in Serum-Containing Media and Serum-Free Media (혈청배지와 무혈청배지에서의 재조합 CHO 세포 성장과 Erythropoietin 생산)

  • 변태호;전복환
    • KSBB Journal
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    • v.11 no.3
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    • pp.288-294
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    • 1996
  • We have investigated the characteristics of recombinant CHO cell growth and erythropoletin(EPO) production at different concentrations of serum and inoculation density. Cell growth and EPO production were increased with the increase of serum concentration and inoculation density. Enhancement of CHO cell growth and EPO production by medium exchange using serum-free medium at the growth phase of cells was studied. It was found that the exchange of culture medium with serum-free medium was favorable for growth of cells and production of EPO. The maximum number of cell and concentration of EPO obtained by exchanging culture medium were $6.2{\times}105cells/$\textrm{cm}^2$ and 7,470units/m1, respectively, compared to $2.1{\times}105cells/\textrm{cm}^2$ and 2,380units/m1 in serum-containing medium without medium exchange. It was observed that CHO cell growth was correlated with EPO production in serum-free media.

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Enhancement of Hybridoma Cell Growth and Anti-Hepatitis B Surface Antigen Monoclonal Antibody Production in Enriched Media with Low Serum (저혈청농축배지에서 세포성장 및 간염표면항원에 대한 단일클론항체 생산의 증가)

  • 전복환;조의철김동일백승복
    • KSBB Journal
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    • v.5 no.1
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    • pp.87-94
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    • 1990
  • Enhancement of hybridoma cell growth and monoclonal antibody(MAb) production by the addition of a small amount of serum into both serum-free medium and enriched medium was studied. The enriched medium was constructed by mixing a basal serum-free medium and a nutrient-fortified RPMI 1640 medium. It was supplemented with human serum albumin, insulin, transferrin, and monoethanolamine. It was found that addition of low concentration of serum with other serum-free supplements was favorable for growth of a mouse hybridoma 2c3.1 cells. The concentration of serum was determined to 0.5%. The maximum cell concentration obtained in this enriched medium supplemented with 0.5% fetal bovine serum (FBS) was $3.06{\times}10^6$ cells/ml and the concentration of secreted anti-Hepatitis surface antigen (antiHBsAg) MAb was $159.7{\mu\textrm{g}}\;/\;ml$ compared to $43{\mu\textrm{g}}\;/\;ml$ in RPMI 1640 medium with 10% FBS and $50{\mu\textrm{g}}\;/\;ml$ in previously-developed serum-free medium. The 2c3.1 cell growth and MAb production could be enhanced considerably by using the enriched medium supplemented with 0.5% FBS and serum-free supplements instead of RPMI 1640 medium or serum-free medium. The enhancement in MAb production in the enriched medium was more noticeable.

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Culture of Human Articular Chondrocytes in Serum-free Media

  • Choi, Yong-Soo;Lim, Sang-Min;Lee, Chang-Woo;Kim, Dong-Il
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.335-339
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    • 2003
  • The aim of this study is to optimize the monolayer cultivation of human articular chondrocytes in serum-free media. For this purpose, chondrocytes were isolated from human articular cartilage and monolayer cultures were performed in DMEM/F12 medium with 10% fetal bovine serum (FBS) or serum-free media (SFM) containing various supplements and epidermal growth factor (EGF). Western blotting analysis, RT-PCR, dimethylmethylene blue (DMB) assay were carried out to evaluate the synthesis of collagen type II (Col. II) and glycosaminoglycans (GAGs). We observed that SFM with EGF stimulated the cell growth while the amounts of synthesized GAGs and Col. II were decreased gradually. However, the Col. II mRNA level was increased when the SFM was replaced by media containing 10% FBS. This study suggests that it is possible to obtain large amount of human articular chondrocytes by short-term monolayer cultures in SFM.

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Lipids Supply for Hybridoma Culture (하이브리도마 배양을 위한 lipids의 공급)

  • 박상재;최차용
    • KSBB Journal
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    • v.8 no.5
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    • pp.483-487
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    • 1993
  • BSA/acids component in serum free medium (SFM) developed for the culture of hybridoma cell line, KA112, was replaced by acids/Pluronic F-68 emulsion. Protein content of SFM was minimized, and increased maximum cell density was obtained in serum-free lipids supplemented medium (SFLSM). Cell growth promotion effect of the emulsion was not affected by filtration with 0.2$\mu$m filter.

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Development of Serum-free Media for the Culture of Mouse Hybridoma (I) ; Determination of Optimal Media Composition (쥐 하이브리도마 세포배양을 위한 무혈청 배지개발( I ) -최적 배지성분의 결정-)

  • 조보연;최태부
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.481-488
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    • 1989
  • A serum-free medium that could be used for the large-scale culture of mouse hybridoma to produce monoclonal antibodies was developed. The medium was based on a 1:1 mixture of Iscove's Modified Dulbecco's Medium and Ham's F-12, supplemented with insulin 10$\mu\textrm{g}$/$m\ell$, transferrin 10$\mu\textrm{g}$/$m\ell$, ethanolamine 10$\mu$M and selenium 30nM (designated EBM (enriched basal medium) with the supplements). The effect of various supplements of steroid hormones, vitamins, lipid and mineral salts was investigated and their optimal concentration was determined to replace fetal calf serum (PCS). These components were added respectively and then added by way of two or three combination to discern of which component combination was effective to the culture of hybridoma. As a result, serum-free medium KM3 (EBM with BSA 100$\mu\textrm{g}$/$m\ell$, mineral cocktail and 0.05% PEG) was deter-mined. The hybridoma Alps 25-3 cultured in this medium showed almost the same growth rate as in medium added with 2% fetal bovine serum. However, the antibody concentration from KM3 cultures was 80% of that obtained from culture with FCS. KM3 was also examined for the culture of other mouse hybridomas, KW, A4W & HCGK, and it was confirmed that it could support the growth of these hybridomas and the production of monoclonal antibodies.

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Suspension culture of anchorage-dependent cells in serum-free medium with biodegradable polymer nanospheres

  • Ryu, Ju-Hee;Choi, Cha-Yong;Kim, Byung-Soo
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.171-173
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    • 2003
  • Suspension culture in serum-free medium is important for the efficient large-scale culture of anchorage-dependent cells that are utilized to produce therapeutic recombinant protein(e.g., insulin, antibody, vaccine) and virus vector for therapeutic gene transfer. We developed a novel method for the suspension culture of anchorage-dependent animal cells in serum-free medium using biodegradable polymer nanospheres in this study. Poly(lactic-co-glycolic acid) (PLGA) polymer nanospheres (433nm in average diameter) were used to the culture of human embryonic kidney 293 cells in serum-free medium in stirred suspension bioreactors. The use of PLGA nanospheres promoted the aggregate formation and cell growth (3.8-fold versus 1.8-fold growth), compared to culture without nanospheres. Adaptation of the anchorage-dependent cells to suspension culture or serum-free medium is time-consuming and costly. In contrast, the culture method developed in our study does not require the adaptation process. This method may be useful for the large-scale suspension culture of various types of anchorage-dependent animal cells in serum-free medium.

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Development of Serum-Free Medium for Mouse-mouse Hybridoma Part II. Hybridoma Culture using Developed Serum-Free Media (Hybridoma배양을 위한 무혈청 배지의 개발 제2부 : 무혈청 배지를 사용한 Hybridoma배양)

  • 제훈성;최차용
    • KSBB Journal
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    • v.8 no.1
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    • pp.28-35
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    • 1993
  • The serum free medium was developed and used for the suspension culture of mammalian cells. Although there were the problems of the longer lag time and the smaller maximum cell concentration achievable, the higher specific productivity as well as other advantages of the serum free medium can make it a more realistic alternative. The existence of a staggering period in glucose concentration vs. time profile in the batch culture can be a practical indicating signal for performing fed batch culture. The concentration dependence of the effects of the additives in the serum free medium as well as its economic feasibility was also tested.

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