• Title/Summary/Keyword: 3D cell culture

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Light-susceptibility of Camptothecin Production from in Vitro Cultures of Camptotheca acuminata Decne

  • Park, Young-Goo;Park, Mee-Hee;Yang, Jae-Kyung;Chung, Young-Gwan;Park, Myung-Suk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.1
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    • pp.32-36
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    • 2003
  • Production of camptothecin ((PT) from callus cultures of Camptotheca acuminata Decne was affected by light and culture conditions. Among the culture media tested, modified B5 medium containing 3% (w/v) sucrose, 2 mg/L B,4-D, 2 times of MS medium vitamins, 500 mg/L casein hydrolysate, 250 mg/L myo-inositol, 0.05% (w/v) activated charcoal, and 0.15% (w/v) gelite was used for callus induction . The highest cell growth and CPT production were obtained in dark and green light condition, respectively. Photoperiod has no effect on cell growth and CPT production. Both cell growth and CPT production were also influenced by combination ratio of red and blue light .Cell growth and CPT production were the highest in the ratio of red and blue light,90:10.

Direct Fermentation of D-Xylose to Ethanol by Candida sp. BT001

  • LEE, SANG-HYEOB;WON-GI BANG
    • Journal of Microbiology and Biotechnology
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    • v.4 no.1
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    • pp.56-62
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    • 1994
  • A yeast strain, BT001, which can directly ferment D-xylose to ethanol was isolated from forest soils, and then identified as Candida sp. Cultural conditions for the optimum ethanol production, along with the effects of aeration on cell growth and ethanol production were investigated. Aeration stimulated the cell growth and the volumetric rate of ethanol production, but decreased the ethanol yield. Optimum temperature and initial pH for the ethanol production were $33{\circ}^C$ and 6.0, respectively. In a shake flask culture, this strain produced 52.3 g ethanol per liter from 12%(w/v) D-xylose after incubation for 96 hours. Ethanol yield was 0.436 g per g D-xylose consumed. This corresponds to 85.8% of theoretical yield. Also, this yeast strain produced ethanol from D-galactose, D-glucose and D-mannose, but not from L-arabinose and L-rhamnose. Among these sugars, D-glucose was the fastest in being converted to ethanol sugars.

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Effect of Energy Sources (Glucose, Pyruvate and Lactate) Added to Dulbecco's Modified Eagle Medium (DMEM) on the Mouse 2-cell Embryo Development (Glutamine 함유 배양액에 첨가한 에너지원이 마우스의 배 발달에 미치는 영향)

  • Kim, Ju-Hwan;Park, Kee-Sang;Lee, Taek-Hoo;Chun, Sang-Sik;Song, Hai-Bum
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.1
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    • pp.1-7
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    • 2000
  • Objective: Mammalian embryos undergo changes of energy environment for transfer from oviduct to uterus. Also, the human reproductive organ (oviduct, uterus) contains energy sources of different concentration (oviduct - glucose: 0.5 mM, pyruvate: 0.32 mM, lactate: 10.5 mM; uterus - goucose: 3.15 mM, pyruvate: 0.1mM, lactate: 5.87 mM, respectively). This study was conducted to examine the effect of these energy sources added in DMEM with glutamine on the mouse embryo development. Methods: There was used ICR female mouse. Two cell embryos of mouse are collected by method of 'flushing'. Flushing fluid was used Ham's F-10 added to 20% FBS. The collected 2 cell embryos were cultured in media such as Control (only DMEM), group A and B (DMEM supplemented with 0.5 mM and 3.15 mM glucose), and group C and D (DMEM supplemented with 0.1 mM and 0.32 mM pyruvate), and group E and F (DMEM supplemented with 5.87 mM and 10.5 mM lactate). All experimental media supplemented with 20% hFF, respectively. Pattern of embryo development was observed to interval at 24hr during 96hr. Results : The media with glutamine added glucose (group A: 51.0%; group B: 48.4%) was significantly (p<0.05) higher than other experimental group in development into the morula stage after 24 hr in culture, but not significantly different compared with control and the rate of development into the blastocyst was significantly (p<0.05) low in the both of pyruvate (group C: 7.9% group D: 6.8%) and lactate (group E: 7.1%, group F: 7.1%) treatment group after 48 hr in culture. Development into the blastocyst and hatched balstocyst after 72 hr in culture revealed similarly in control (81.9%) and glucose treatment group (group A: 83.3%, group B: 82.8%). However, development into the hatched and attached blastocyst after 96hr in culture revealed significantly (p<0.05) development in the glucose treatment group (group A: 82.3%, group B: 78.5%) than control (63.2%), and its of pyruvate (group C: 34.1%, group D: 34.1%) and lactate (group E: 25.9%, group F: 33.3%) treatment group were significantly (p<0.05) lower than control similar to previous observations. Conclusion : The glucose added to the DMEM with only glutamine, as energy source, was highly to the rate of development compared with control, but the other energy sources were not, synthetically. Above refer to, the human reproductive organ (oviduct, uterus) contains energy sources of different concentration. Thus, further studies are will examine continuously to effects by interaction of different energy sources in the mouse embryo development, and these results will provide to foundation on the human embryo culture.

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Heterocyclic Amines Removal by Binding Ability of Lactic Acid Bacteria Isolated from Soybean Paste (된장에서 분리된 유산균의 결합력에 의한 Heterocyclic Amines 제거)

  • Lim, Sung-Mee
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.73-83
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    • 2014
  • The objective of the this study was to investigate the binding capacity and removal ability of lactic acid bacterial strains obtained from Korean soybean paste for mutagenic heterocyclic amines (HCAs) formed during cooking of protein-rich food at high temperature. Among 19 strains identified by carbohydrate fermentation and 16S rRNA sequencing, the live cell or cell-free culture supernatant of Lactobacillus acidophilus D11, Enterococcus faecium D12, Pediococcus acidilactici D19, L. acidophilus D38, Lactobacillus sakei D44, Enterococcus faecalis D66, and Lactobacillus plantarum D70 inhibited the mutagenesis caused by either 3-amino-1,4-dimethyl-5H-pyrido[4,3-b] indole (Trp-P-1) or 3-amino-1-methyl-5H-pyrido[4,3-b] indole (Trp-P-2) in Salmonella typhimurium TA98 and TA100. The bacterial cells of the isolated strains showed greater binding activity than the pure cell wall, exopolysaccharide, and pepetidoglycan. The carbohydrate moieties of the cell wall or protein molecules on the cell surface have a significant role in binding Trp-P-1 and Trp-P-2, since protease, heating, sodium metaperiodate, or acidic pH treatments significantly (P<0.05) reduced the binding efficacy of the tested bacteria. Addition of metal ions or sodium dodecyl sulfate decreased the binding ability of E. faecium D12, L. acidophilus D38, and E. faecalis D66. Therefore, the binding mechanisms of these strains may consist of ion-exchange and hydrophobic bonds. Especially, the high mutagen binding by L. acidophilus D38 and L. plantarum D70 may reduce the accumulation or absorption of Trp-P-1 and Trp-P-2 in the small intestine via increased excretion of a mutagen-bacteria complex.

Optical Microscope Image Processing for Automated Cells Counting (세포 자동 계수를 위한 광학현미경 이미지 처리)

  • Cho, Mi-Gyung;Moon, Sang-Jun;Shim, Jae-Sool
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.15 no.11
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    • pp.2493-2499
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    • 2011
  • With growth of nano-bio industry, it is of significant importance to develop an automated system to exploit cell behaviors, including migration, mitosis, apoptosis, shape deformation of individual cells and their interactions among cells in the process of cell growth. In this paper, we proposed preprocessing techniques, a classification method which classifies clusters (overlapping multiple cells) from cells and an automated method which counts the number of cells and clusters in order to analyze 2D or 3D deformations of the cells in the real-time images from microscope in the cell culture. We conducted the 3T3 cell images taken from each thirty-minute interval. It showed the average 99.8% accuracy automatically for separating cells and clusters.

Selection of Mutant Silkworm with Oxidation-deficient Haemolymph for Insect Cell Culture (곤충세포 배지 개발을 위한 체액산화지연 돌연변이 누에계통 선발)

  • Choi, Ji-Young;Kim, Jong-Gill;Choi, Young-Cheol;Yoon, Hyung-Joo;Ahn, Mi-Young;Kim, Sam-Eun;Hwang, Seok-Jo
    • Journal of Sericultural and Entomological Science
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    • v.49 no.2
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    • pp.47-50
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    • 2007
  • Insect cell culture system has been demonstrated the effective means of producing medical and agricultural products. Furthermore, Fetal bovine serum (FBS) is in wide use in insect cell culture. Silkworm hemolymph was tested to develop as a substitute for FBS and was effective in insect cell growth. Hemolymph is oxidized and darkens visibly during the collection from silkworms due to the activity of tyrosinase in it. Toxic quinones are produced by the oxidation and consequently inhibit the cell growth. Heat treatment can be used to prevent the oxidation; however, the oxidation may occur during the collection of hemolymph before it is heat-treated. Hemolymphs collected from 257 different strains of silkworms were examined to select the slowly oxidized hemolymphs. Hemolymphs collected from mutant strains such as $Y_4$, TBO and $wE^b$ showed relatively slow color changes. Oxidation rates of the hemolymphs were measured by the absorbance change using a spectrophotometer. The absorbance of mutant hemolymph reached the saturation value at $20^{\circ}C$ in each 330 min ($Y_4$), 360 min (TBO) and 450 min ($wE^b$) min, whereas the total oxidation time of the wild-type (Baekokjam) hemolymph at the same temperature was 120 min. The cell growth in the medium supplemented with mutant species hemolmph was more effective that in the medium supplemented with Baekokjam species hemolymph.

Electrohydrodynamic Jet Process for Pore-Structure-Controlled 3D Fibrous Architecture As a Tissue Regenerative Material: Fabrication and Cellular Activities

  • Kim, Minseong;Lee, Hyeongjin;Kim, GeunHyung
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2017.05a
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    • pp.134.1-134.1
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    • 2017
  • In this study, we propose a new scaffold fabrication method, "direct electro-hydrodynamic jet process," using the initial jet of an electrospinning process and ethanol media as a target. The fabricated threedimensional (3D) fibrous structure was configured with multilayered microsized struts consisting of randomly entangled micro/nanofibrous architecture, similar to that of native extracellular matrixes. The fabrication of the structure was highly dependent on various processing parameters, such as the surface tension of the target media, and the flow rate and weight fraction of the polymer solution. As a tissue regenerative material, the 3D fibrous scaffold was cultured with preosteoblasts to observe the initial cellular activities in comparison with a solid-freeform fabricated 3D scaffold sharing a similar structural geometry. The cell-culture results showed that the newly developed scaffold provided outstanding microcellular environmental conditions to the seeded cells (about 3.5-fold better initial cell attachment and 2.1-fold better cell proliferation).

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Factors influencing efficiency of somatic embryogenesis of Gentiana kurroo (Royle) cell suspension

  • Fiuk, Agnieszka;Rybczynski, Jan J.
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.33-39
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    • 2008
  • In this paper, we would like to show unexpected morphogenic potential of cell suspensions derived from seedling explants of Gentiana kurroo (Royle). Suspension cultures were established with the use of embryogenic callus derived from seedling explants (root, hypocotyl and cotyledons). Proembryogenic mass proliferated in liquid MS medium supplemented with $0.5mg\;l^{-1}$ 2,4-D and $1.0mg\;l^{-1}$ Kin. The highest growth coefficient was achieved for root derived cell suspensions. The microscopic analysis showed differences in aggregate structure depending on their size. To assess the embryogenic capability of the particular culture, 100 mg of cell aggregates was implanted on MS agar medium supplemented with Kin ($0.0-2.0mg\;l^{-1}$), $GA_3$ ($0.0-2.0mg\;l^{-1}$) and AS ($80.0mg\;l^{-1}$). The highest number of somatic embryos was obtained for cotyledon-derived cell suspension on $GA_3$-free medium, but the best morphological quality of embryos was observed in the presence of $0.5-1.0mg\;l^{-1}$ Kin, $0.5mg\;l^{-1}$ $GA_3$ and $80.0mg\;l^{-1}$ AS. The morphogenic competence of cultures also depended on the size of the aggregate fraction and was lower when size of aggregates decreased. Flow cytometry analysis reveled luck of uniformity of regenerants derived from hypocotyl suspension and 100% of uniformity for cotyledon suspension.

Plant Regeneration from Cell Suspension Culture Using Leaf Callus in Actinidia deliciosa X A. arguta Clone 118 (양다래X다래 클론 118의 엽조직 캘러스를 이용한 세포 현탁배양으로부터 식물체 유도)

  • Kim Yong-Wook;Moon Heung-Kyu
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.287-292
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    • 2005
  • Calli were induced by culturing the leaf segment of Actinidia deliciosa ${\times}$ A. arguta clone 118 on MS medium supplemented with 0.5 mg/L 2,4-D, 0.1 mg/L NAA and 0.05 mg/L BA for 8 weeks in light condition. The induced calli were inoculated in liquid MS medium containing 0.5 mg/L 2,4-D, 0.1 mg/L NAA, 0.05 mg/L BA and 3% sucrose to establish cell suspension culture. The cells at the exponential stage and the stationary stage could be observed between 5-11 days and after that 12 days in culture, respectively. The fresh weight of callus induced from the suspended cells did not vary much among the media containing eight different combinations of plant growth regulators tested. The highest frequency of shoot induction (88.3%) was observed in MS medium containing 2.0 mg/L zeatin. Either BA or zeatin mixed with thidiazuron (TDZ) seemed to be effective in shoot induction. The induced shoots were transferred to MS medium containing 0.2 mg/L zeatin for further shoot growth. And then the shoots were transferred to Standardi (ST) medium containing 1.0 mg/L indolebutyric acid (IBA) for rooting. Plantlets could be obtained through cell suspension culture of Actinidia deliciosa ${\times}$ A. arguta clone 118.