• Title/Summary/Keyword: high density cell culture

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Studies on the Differentiation of Chondrogenic Cells in Developing Chick Embryo I. Cellular Aggregation and Chondrogenesis (발생계배 연골세포의 분화기구에 대한 연구 I. 세포응집과 분화와의 관계)

  • 박대규;손종경;유정아;유병제;강신성
    • The Korean Journal of Zoology
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    • v.33 no.3
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    • pp.310-321
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    • 1990
  • To establish the in vitro culture system and quantitation for chondrogenesis, and to investigate the relationship between cell aggregation and chondrogenesis, chick limb bud mesenchymal cells of Hamburger-Hamilton stage 23/24 were micromass cultured in various cell densities. The chondrogenesis was assayed based on checking the alcian blue-stained nodule numbers, the amount of alcian blue extraded, the change in cell numbers, the rate of [35 S] sulfate incorporation and expression of type II collagen. Mesenchymal cells plated with an initial density of high (1 x 107 cells/ml)- and intermediates (5. $\times$ 106 cells/ml)-density were differentiated into cartilage. On the other hand, the cells of low density (2 x 106 cells/mi, 5 $\times$ 105 cells/ml) of stage 23/24 cells and the stage 18/19 cells in three kinds of cell density did not differentiate into cartilage even though the cells formed an aggregated core at the center of cultured mass. From these results and others obtained in this study, it can be stated that the stage 23/24 mesenchymal cells are likely to pass over the aggregation step and have the potentiality to differentiate into chondrocytes. Thus chondrogenesis in vitro can be observed when mesenchymal cells are plated over the threshold density of 5 $\times$ 106 cells/ml. Hyaluronidase (HAase) activity was relatively constant throughout the culture, suggesting that the role of HAase may not be important for the cells of stage 23/24.

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Development of Curdlan Separation Process with Density Gradient Centrfugation (Density Gradient를 이용한 식품소재를 커들란의 분리공정개발)

  • 김봉영;이중헌
    • KSBB Journal
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    • v.16 no.5
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    • pp.523-525
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    • 2001
  • Curdlan is one biopolymer composed of ${\beta}$1,3-glucan and dissolved in a alkali solution but formed salt under neutral or acid condition. It was produced by Agrobactrium species and the separation process is necessary to make pure curdlan from the culture broth. The pH swing separation method was as feasible separation process using solubility changes with the pH difference. however, this method requires a lot of acid and alkali solution also produces a lot of waste. Therefore, an efficient process which could save energy and minimize toxic waste was developed. A density gradient separation process was developed in this research. High density sucrose solution was used as a separation agent. Curdlan was separated from the culture broth when the density of the sucrose solution was 1.15 g/L. Since the curdlan was produced on the surface of cell wall. the pre-treatment of culture broth was necessary. Curdlan recovery yield was increased up to 83% with the homogenization of the culture broth and further increased up to 87% with the treatment of alkai-acid solution.

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Characteristics of Immobilized Culture of Mentha piperita Cells for Oil Production

  • Ha, Won Ho;Gun Jo Woo;Hyong Joo Lee
    • Journal of Microbiology and Biotechnology
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    • v.6 no.2
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    • pp.132-136
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    • 1996
  • To investigate the characteristics of immobilized peppermint (Mentha piperita) cells, dry cell weight (DCW), change of cell viability, and oil productivity of the immobilized cells were determined. Peppermint cells were immobilized in polyurethane (PU) foams of $5{\times}5{\times}5$ mm and cultured in a shaking flask. The maximum DCW was 2.1 mg per foam piece after 20 days of cultivation and the cell density was approximately 420 mg per flask containing 200 foams in 200 ml medium. For the first five days of cultivation, the cell viability was about 80$%$ and decreased to 70$%$ during 5 to 20 days of cultivation. The maximum oil productivity, 148 mg/l was achieved after 40 days of cultivation. The immobilized cells were also cultivated in a bioreactor, equipped with a round spiral type impeller, containing 2, 400 PU foams. The cell viability after 30 days of cultivation with chitosan as an elicitor in the bioreactor was 67$%$ and DCW was 2.0 mg per foam piece. Though the cell viability was relatively high in the bioreactor system, the oil productivity was relatively lower than that of the flask system.

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Production of Pigment by Liquid Culture and Monacolin K in Red Mold Rice by Solid State Fermentation of Monascus ruber Strains (Monascus ruber의 액체배양을 통한 색소 생산 및 고체발효를 통한 홍국쌀의 monacolin K 생산 특성)

  • Park, Youn-Je
    • KSBB Journal
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    • v.28 no.6
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    • pp.400-407
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    • 2013
  • The growth characteristics and production of color pigments by Monascus strains were investigated during liquid culture, and production of monacolin K in red mold rice was carried out by solid state fermentation. Four different Monascus ruber strains were cultured in potato dextrose yeast extract broth (PDYB) media at $25^{\circ}C$ for 15 days. The high producing strain for red pigment was not corresponded to the strain for yellow pigment. Production of red pigment was high in the strain causing the fast pH change in culture broth. Production of monacolin K in red mold rice by solid state fermentation was influenced by a combination of wet cell weight and spore density in inoculum by liquid culture. Most strains showed the high production of monacolin K in red mold rice, when submerged fermentation was carried out for 5 days as inoculum for solid state fermentation. These results suggest that submerged fermentation period of inoculum have an effect on the production of monacolin K in red mold rice by solid state fermentation, and monacolin K in red mold rice could be increased by controlling the condition of submerged fermentation for inoculum.

THE MOST APPROPRIATE ANTIMITOTIC TREATMENT OF ARA-C IN SCHWANN CELL-ENRICHED CULTURE FROM DORSAL ROOT GANGLIA OF NEW BORN RAT (신생 백서 척수후근절의 슈반세포 배양을 위한 Ara-C 분열억제제의 최적 효과에 대한 연구)

  • Kim, Soung-Min;Lee, Jong-Ho;Ahn, Kang-Min;Kim, Nam-Yeol;Sung, Mi-Ae;Hwang, Soon-Jeong;Kim, Ji-Hyuck;Jahng, Jeong-Won
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.2
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    • pp.100-107
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    • 2004
  • Schwann cell, one of important components of peripheral nervous system, interact with neurons to mutually support the growth and replication of embryonal nerves and to maintain the different functions of adult nerves. The Ara-C, known as an antimitotic agent, have been used to have high effectiveness in eliminating fibroblasts during Schwann cell culture period. This enrichment effect is also known to be cummulative with each successive pulse of Ara-C applied and is due to a progressive loss of fibroblasts. But the cytotoxicity by Ara-C is also cummulative and noticeable over the period. To determine the most effective application time and interval of Ara-C in the Schwann cell culture, we observed the Schwann cell purity and density with the Ara-C treatment in plain and three-dimensional culture from dorsal root ganglion of new born rat. By culturing dispersed dorsal root ganglia, we can repeatedly generate homogenous Schwann cells, and cellular morphology and cell count with mean percentages were evaluated in the plain culture dishes and in the immunostainings of S-100 and GFAP in the three-dimensional culture. The Ara-C treated cultures showed a higher Schwann cell percentage ($31.0%{\pm}8.09%$ in P4 group to $65.5%{\pm}24.08%$ in P2 group), compared with that obtained in the abscence of Ara-C ($17.6%{\pm}6.03%$) in the plain culture after 2 weeks. And in the three-dimensional culture, S-100 positive cells increased to $56.22%{\pm}0.67%$ and GFAP positive cells to $66.46%{\pm}1.83%$ in G2 group (p<0.05), higher yield than other groups with Ara-C application. Therefore, we concluded that the Ara-C treatment is effective for the proliferation of Schwann cells contrast to the fibroblasts in vitro culture, and the first application after 24 hours from cell harvesting and subsequent 2 pulse treatment (P2 group in plain culture and G2 group in three-dimensional culture) was more effective than other application protocols.

The most appropriate antimitotic treatment of Ara-C in Schwann cell-enriched culture from dorsal root ganglia of new born rat

  • Kim, Soung-Min;Jahng, Jeong-Won;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.1
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    • pp.42-51
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    • 2006
  • Schwann cell, one of important components of peripheral nervous system, interact with neurons to mutually support the growth and replication of embryonal nerves and to maintain the different functions of adult nerves. The Ara-C, known as an antimitotic agent, have been used to have high effectiveness in eliminating fibroblasts during Schwann cell culture period. This enrichment effect is also known to be cummulative with each successive pulse of Ara-C applied and is due to a progressive loss of fibroblasts. But the cytotoxicity by Ara-C is also cummulative and noticeable over the period. To determine the most effective application time and interval of Ara-C in the Schwann cell culture, we observed the Schwann cell purity and density with the Ara-C treatment in plain and three-dimensional culture from dorsal root ganglion of new born rat. By culturing dispersed dorsal root ganglia, we can repeatedly generate homogenous Schwann cells, and cellular morphology and cell count with mean percentages were evaluated in the plain culture dishes and in the immunostainings of S-100 and GFAP in the three-dimensional culture. The Ara-C treated cultures showed a higher Schwann cell percentage (31.0%${\pm}$8.09% in P4 group to 65.5%${\pm}$24.08% in P2 group), compared with that obtained in the abscence of Ara-C (17.6%${\pm}$6.03%) in the plain culture after 2 weeks. And in the three-dimensional culture, S-100 positive cells increased to 56.22%${\pm}$0.67% and GFAP positive cells to 66.46%${\pm}$1.83% in G2 group (p<0.05), higher yield than other groups with Ara-C application. Therefore, we concluded that the Ara-C treatment is effective for the proliferation of Schwann cells contrast to the fibroblasts in vitro culture, and the first application after 24 hours from cell harvesting and subsequent 2 pulse treatment (P2 group in plain culture and G2 group in three-dimensional culture) was more effective than other application protocols.

Enhanced reutilization value of shrimp-shell waste via fed-batch biodegradation with higher production of reducing sugar, antioxidant, and DNA protective compounds

  • Rashid, Harun Ar;Jung, Hyun Yi;Kim, Joong Kyun
    • Fisheries and Aquatic Sciences
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    • v.21 no.10
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    • pp.33.1-33.11
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    • 2018
  • As a process for commercial application, production of reducing sugar, antioxidant, and DNA protective compounds from shrimp-shell powder was investigated in a fed-batch biodegradation using Bacillus cereus EW5. The fed-batch biodegradation was operated in a 5-L bioreactor for 96 h according to three times pulse-feeding strategy. On the basis of the equal working volume (3 L), the fed-batch biodegradation showed a better production of the target compounds than the batch biodegradation, with higher cell density and shortened biodegradation period. The maximum values of the target compounds were 0.297 mg/mL of reducing sugar, 92.35% DPPH radical scavenging activity, 98.16% ABTS radical scavenging activity, and 1.55 reducing power at $A_{700}$, which were approximately 12.1, 3.4, 5.2, and 8.4% enhanced, respectively, compared with those obtained from the batch biodegradation. The fed-batch culture supernatant also showed the enhanced DNA damage inhibition activity than the batch culture supernatant. As a result, the fed-batch biodegradation accompanied by high cell density could produce more useful compounds, enabling an increase in the reutilization value of shrimp-shell waste.

Effects of Cell Size, Density, Conditioned Media and pH on Carrot (Daucus carota L.) Cell Embryogenesis (당근(Daucus carota L.) 세포 배양시 세포의 크기, 밀도, Conditioned 배지 및 pH가 배발생에 미치는 영향)

  • 백기엽;이철희;황주광
    • Journal of Plant Biology
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    • v.28 no.2
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    • pp.141-148
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    • 1985
  • The effects of sizes and densities of cells cultured, conditioned medium, and media pH on the somatic embryogenesis of carrot (Daucus carota L.) were examined. A large number of globular embryoids was formed after 4 days in cell culture, and later globular embryoids developed into heart and torpedo shape. High cell density resulted in higher number and better growth of embryos, especially on conditioned medium than Murashige-Skoog medium. The fresh weight and number of embryoids formed increased with the decrease in cell size. The significant reduction in fresh weight and number of embryoids was obtained when culturing cells with diameter of over 90 ${\mu}{\textrm}{m}$. Dry weight and number of embryoids were markedly reduced with medium pH of 4 or 7, but promoted with pH 6.0.

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Enhancement of Biocontrol Activity of Serratia plymuthica A21 -4 Toward Phytophthora Blight of Pepper by Amendment of Nutritional Condition

  • Shen, Shun-Shan;Kim, Chang-Guk;Park, Chang-Seuk
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.96.1-96
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    • 2003
  • Serratia plymuthim A21-4 strongly inhibits the mycelial growth, zoospore formation, and cystospore germination of Phytophthor spp and Pythium species. The bacterial isolate produced antifungal substance and chitinase. The bacteria also enhanced to plant growth remarkably in low nutritional condition. The application of cell suspension of A21-4 to pepper seedlings in greenhouse experiments and soil drenching in farmer's field was proved successfully to control the phythophthora blight of pepper. For the effective control, however, relatively high density of cell number(10$\^$9/cfu/$m\ell$) is required. Density effect was similar in plant growth promoting activity of A21-4. Though this investigation we improved the problem with changes of culture condition of bacteria and some nutritional amendment.

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Effect of Liquid Circulation Velocity and Cell Density on the Growth of Parietochloris incisa in Flat Plate Photobioreactors

  • Changhai Wang;Yingying Sun;Ronglian Xing;Liqin Sun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.2
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    • pp.103-108
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    • 2005
  • For more accurately describing the durations of the light and the dark phases of micro-algal cells over the whole light-dark cycle, and probing into the relationship between the liquid circulation time or velocity, the aeration rate and cell density, a series of experiments was carried out in 10 cm light-path flat plate photobioreactors. The results indicated that the liquid flow in the flat plate photobioreactor could be described by liquid dynamic equations, and a high biomass output, higher content and productivity of arachidonic acid, $70.10\;gm^{-2}d^{-1},\;9.62\%$ and 510.3 mg/L, respectively, were obtained under the optimal culture conditions.