• Title/Summary/Keyword: serum-free media

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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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Development of Serum-Free Media for Primary Culture of Human Articular Chondrocytes

  • CHOI YONG SOO;LIM SANG MIN;LEE CHANG WOO;KIM DONG-IL
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1299-1303
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    • 2005
  • Human articular chondrocytes (HAC) were cultivated as a monolayer in a serum-free medium for primary culture (SFM-P). An optimized SFM-P provides $95\%$ proliferation rate of that obtainable from primary and secondary chondrocyte cultures grown in a control medium with serum. The gradual decrease in the amounts of synthesized glycosaminoglycan and type II collagen was improved by coating the culture dishes with type IV collagen and fibronectin. A significant improvement in the expression of type II collagen and aggrecan mRNA could be achieved. In addition, the monolayer cultures showed better synthesis of the extracellular matrices than alginate-bead cultures in SFM-P.

Medium Recycle Process for the Production of scu-PA from Serum Free Medium (무혈청 배지로부터 scu-PA의 생산을 위한 배지의 재사용 공정에 관한 연구)

  • 김영남;박경유
    • KSBB Journal
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    • v.8 no.1
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    • pp.17-22
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    • 1993
  • $1.96{\times}10^{-5}$(IU/cell/hr) of specific scu-PA production rate was obtained from HEK cells in maintaining ca. $8{\times}10^{5}$(cells/ml) of maximum roll density at 10(ml/hr) of perfusion rate with recycling 20% serum free conditioned media. It can be compared to $4{\times}10^{6}$(cells/ml) of maximum cell density and $4.56{\times}10^{-4}$(IU/cell/hr) of specific production rate in cultivating cells with 1% serum containing medium. Thc conversion ratio of scu-PA to tc-UK increased up to 55% as the recycling ratio increased; however, recycling the used medium seemed to have least negative effect on cell growth. It also showed that the recycling process had definitive advantage of using serum free medium in perfusion cultivation of HEK cell line.

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Optimization of Human Thrombopoietin Production in Insert Cells Using Baculovirus Expression System (베큘로 바이러스 발현 시스템에 의한 곤충세포에서의 인간 트롬보포이에틴 생산 최적화)

  • 고여욱;손미영;박상규;안혜경;박승국;박명환;양재명
    • KSBB Journal
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    • v.13 no.2
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    • pp.181-186
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    • 1998
  • In order to obtain high-level production of recombinant human thrombopoietin (rhTPO) in insect cell line, HTI-TN-5B1-4 (TN5), conditions for optimal rhTPO expression such as multiplicity of infection (MOI), the cell density at infection, harvesting time and type of culture method as well as growth media were determined. When TN5 cells were cultured as anchorage-dependent state in 60-mm dish, cell density $2\times^6$ cells,MOI of 10 and Garvesting the culture media at 72 hr post-infection wrere the cinditions for highest rh TPO production. High production of rhTPO was also achieved by using EXPRESS FIVE serum free media rather than SF900II serum free media-1. Anchorage-dependent TN5 cells were adapted as a suspension culture when they were grown in the presence of heparin. TN5 cells were successfully cultured at 0.2 L scale in suspension culture without having aggregation. When TN5 cells were cultured as suspension state, cell density of $0.6\times10^6$ cells/mL, MOI of 1 and harvesting the culture media at 72 hr post-infection, gave the highest yield of rhTPO.

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Comparative Characterization of Growth and Recombinant Protein Production among Three Insect Cell Lines with Four Kinds of Serum Free media

  • Kwon, Mi-Sun;Takashi Dojima;Park, Enoch Y.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.142-146
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    • 2003
  • Three insect cell lines, Sf9, Sf21 and Tn5Bl-4, and four different kinds of serum free media (SFM), Sf 900 II, EX-CELL 420, EX-CELL 405 and Express Five, were used to compare the nutrient consumption, byproduct formation, production of recombinant protein and protease activity in suspension cultures. The Sf 900 II SFM was a ppropriate for the cell growth and protein production of the Sf9 and Sf21 cell lines. When the Tn5Bl-4 cell line was grown in the Express Five SFM, the specific growth rate was 1.6 fold higher than those of either the Sf9 or Sf21 cell lines. The glucose and glutamine consumption rates per cells, were 4 and 2.3 times higher than those of the Sf9 cell line, respectively. The overall yield coefficients of the lactate and ammoniumion were 2.8 and 1.5 times higher compared to those of the Sf9 cell line. respectively. The maximum specific ${\beta}$-galactosidase production rate was 4.5 fold that of the Sf9 cell line, a 3 times higher protease activity per cell.

Review of the Current Research on Fetal Bovine Serum and the Development of Cultured Meat

  • Lee, Da Young;Lee, Seung Yun;Yun, Seung Hyeon;Jeong, Jae Won;Kim, Jae Hyeon;Kim, Hyun Woo;Choi, Jung Seok;Kim, Gap-Don;Joo, Seon Tea;Choi, Inho;Hur, Sun Jin
    • Food Science of Animal Resources
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    • v.42 no.5
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    • pp.775-799
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    • 2022
  • The purpose of this review is to summarize studies that investigate blood and the main components of fetal bovine serum (FBS) in vertebrates, including major livestock, and review the current research on commercializing cultured meat. Detailed research on FBS is still lacking; however, some studies have shown that FBS consists of proteins, carbohydrates, growth factors, cytokines, fats, vitamins, minerals, hormones, non-protein nitrogen, and inorganic compounds. However, there are few studies on how the composition of FBS differs from blood or serum composition in adult animals, which is probably one of the main reasons for not successfully replacing FBS. Moreover, recent studies on the development of FBS replacers and serum-free media have shown that it is difficult to conclude whether FBS has been completely replaced or serum-free media have been developed successfully. Our review of the industrialization of cultured meat reveals that many basic studies on the development of cultured meat have been conducted, but it is assumed that the study to reduce or replace ingredients derived from fetuses such as FBS has not yet been actively developed. Therefore, developing inexpensive and edible media is necessary for the successful industrialization of cultured meat.

Ammonium Ion Effects and Its In Situ Removal by Using Immobilized Adsorbent in Hybridoma Cell Culture (하이브리도마 세포배양에서 암모늄 이온의 영향 및 고정화 흡착제에 의한 암모늄 이온의 동시제거)

  • 정연호;이해익
    • KSBB Journal
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    • v.11 no.3
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    • pp.329-339
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    • 1996
  • The effects of ammonium ion on cell growth kinetics, monoclonal antibody productivity, and cell metabolism of hybridoma cells were investigated. The mouse-mouse hybridoma cell line VlIIH-8 producing mouse IgG2a was used as a model system. Ammonium ion showed an inhibitory effect on cell growth and monoclonal antibody production. New immobilized adsorbents were developed for the reduction of the inhibitory effect of ammonium ion. The ammonium ion selective zeolite, Phillipsite-Gismondine was entrapped in calcium alginate bead or in dialysis membrane and applied to the hybridoma cell culture system for the in situ removal of ammonium ion from culture media. The effects of ammonium the both serum supplemented and serum free media on the cell growth were studied by applying immobilized adsorbents of calcium alginate bead type. The results demonstrated a substantial enhancement in cell growth. Applying immobilized adsorbents of dialysis membrane type to serum supplemented media also resulted in the stimulation of cell growth, cell viability and monoclonal antibody production.

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Effect of ${\beta}-Estradiol$ on the Growth of Primary Rabbit Proximal Tubule Cells in Serum-free Medium (${\beta}-Estradiol$이 토끼 근위 세뇨관 상피세포의 성장에 미치는 영향)

  • Park, Sang-Ho;Chung, Joo-Ho;Ko, Kye-Chang;Jung, Jee-Chang
    • The Korean Journal of Pharmacology
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    • v.29 no.1
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    • pp.73-83
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    • 1993
  • In order to examine the effect of ${\beta}-estradiol$ on the cell growth, using a primary rabbit kidney poximal tubule cell culture system. We investigated the effect of ${\beta}-estradiol$ on alpha 1 (IV) collagen and ${\beta}-actin$ mRNA levels from primary rabbit kidney cell cultures, and also the effects of 3 growth factors and ${\beta}-estradiol$ supplementation on the growth of primary rabbit kidney proximal tubule cells in the serum-free medium. 1 nM of ${\beta}-estradiol$ showed a sizable potentiation effect on the growth of the proximal tubule cell in serum-free medium, but higher concentration (> 10 nM) of estradiol indeed inhibited the growth. In the absence of hydrocortisone as a growth supplement in serum-free medium, ${\beta}-estradiol$ caused to potentiate the growth of the cell. In the presence of hydrocortisone, ${\beta}-estradiol$ also potentiated the growth of the proximal tubule cells. According to the Northern analysis, ${\beta}-estradiol$ increased the level of ${\beta}-actin$ mRNA, although mRNA level of the alpha I(IV) collagen was not changed significantly.

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Growth and Differentation of Rat Mammary Epithelial Cells Cultured in Serum-free Medium

  • Kim, Dong-Yeum;Jhun, Byung-Hak;Lee, Kyung-Hee;Hong, Seung-Chul;Clifton, Kelly-H.;Kim, Nam-Deuk
    • Archives of Pharmacal Research
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    • v.20 no.4
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    • pp.297-305
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    • 1997
  • A new serum-free defined medium was developed that supports the growth of normal rat mammary epithelial cells. Mammary organoids from the glands of female F344 rats were cultured in a serum-free medium. Monolayer culture colonies developed within a week and remained viable for months in culture. Upon subculture of one-week-old primary colonies, almost the same morphology of colonies was developed. The scrape loading/dye transfer technique showed that most of colonies that developed in a serum-free medium containing EGF, human transferrin, insulin, and hydrocortisone (basal serum-free medium, BSFM) failed to show cell-cell communication. However, colonies cultured in BSFM supplemented with prolactin, $E_2$, and progesterone (complete hormone serum-free medium, CHSFM) showed cell-cell communication at 14 days of primary culture or of subculture. By flow cytometry with FITCPNA and PE-anti-Thy-1.1 monoclonal antibody, we distinguished four RMEC subpopulations in cultures in both media: Thy-1.1+ cells, PNA+ cells, cells negative to both reagents and cells positive to both reagents. It is likely that combined prolactin, cortisol, and insulin in CHSFM stimulate terminal differentiation of clonogenic cells.

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