• Title/Summary/Keyword: culturing media

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Growth and nutrient removal of Chlorella vulgaris in ammonia-reduced raw and anaerobically-digested piggery wastewaters

  • Kwon, Gyutae;Nam, Ji-Hyun;Kim, Dong-Min;Song, Chulwoo;Jahng, Deokjin
    • Environmental Engineering Research
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    • v.25 no.2
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    • pp.135-146
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    • 2020
  • This study was aimed to investigate the possibility of using raw and anaerobically-digested piggery wastewater as culture media for a green microalga Chlorella vulgaris (C. vulgaris). Due to high concentration of ammonia and dark color, the microalga did not grow well in this wastewater. In order to solve this problem, air stripping and NaOCl-treatment were applied to reduce the concentration of NH3-N and the color intensity from the wastewater. Algal growth was monitored in terms of specific growth rate, biomass productivity, and nutrient removal efficiency. As a result, C. vulgaris grew without any sign of inhibition in air-stripped and 10-folds diluted anaerobically-digested piggery wastewater with enhanced biomass productivity of 0.57 g/L·d and nutrient removal of 98.7-99.8% for NH3-N and 41.0-62.5% for total phosphorus. However, NaOCl-treatment showed no significant effect on growth of C. vulgaris, although dark color was removed greatly. Interestingly, despite that the soluble organic concentration after air stripping was still high, the biomass productivity was 4.4 times higher than BG-11. Moreover, air stripping was identically effective for raw piggery wastewater as for anaerobic digestate. Therefore, it was concluded that air stripping was a very effective method for culturing microalgae and removing nutrients from raw and anaerobically-digested piggery wastewaters.

Effects of IL-3 and SCF on Histamine Production Kinetics and Cell Phenotype in Rat Bone Marrow-derived Mast Cells

  • Lee, Haneul Nari;Kim, Chul Hwan;Song, Gwan Gyu;Cho, Sung-Weon
    • IMMUNE NETWORK
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    • v.10 no.1
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    • pp.15-25
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    • 2010
  • Background: Rat mast cells were regarded as a good model for mast cell function in immune response. Methods: Rat bone marrow mast cells (BMMC) were prepared both by recombinant rat IL-3 (rrIL-3) and by recombinant mouse stem cell factor (rmSCF), and investigated for both proliferation and differentiation in time course. Rat BMMC was induced by culture of rat bone marrow cells (BMCs) in the presence of both rrIL-3 (5 ng/ml) and rmSCF (5 ng/ml). Culture media were changed 2 times per week with the cell number condition of $5{\times}10^4/ml$ in 6 well plate. Proliferation was analyzed by cell number and cell counting kit-8 (CCK-8) and differentiation was by rat mast cell protease (RMCP) II and histamine. Results: Cell proliferation rates reached a maximum at 8 or 11 days of culture and decreased thereafter. However, both RMCP II production and histamine synthesis peaked after 11 days of culture. By real time RT-PCR, the level of histidine decarboxylase mRNA was more than 500 times higher on culture day 11 than on culture day 5. By transmission electron microscopy, the cells were heterogeneous in size and contained cytoplasmic granules. Using gated flow cytometry, we showed that cultured BMCs expressed high levels of $Fc{\varepsilon}RI$ and the mast cell antigen, ganglioside, on culture day 11. Conclusion: These results indicate that rat BMMCs were generated by culturing BMCs in the presence of rrII-3 and rmSCF and that the BMMCs have the characteristics of mucosal mast cells.

Biodegradation of Diesel Oil by Microorganisms Isolated from Petroleum Contaminated Site (유류 오염지역으로부터 분리된 균주를 이용만 디젤유의 분해)

  • 박천보;허병기;윤현식
    • KSBB Journal
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    • v.16 no.6
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    • pp.632-637
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    • 2001
  • The cells obtained from diesel contaminated site were tested for diesel degradation by culturing them on the culture medium that contained diesel as the only carbon source. Two strains that grew well in the culture media were separated: one formed white colony and another strain formed yellow colony. When they were cultured together, much higher diesel degradation was obtained compares to that of individual cell culture. Mixed culture of white and yellow colony forming strains grew well with 1%(v/v) diesel and the addition of growth nutrients increased the diesel degradation. Additional nitrogen source was efficient for higher diesel degradation (over 90%) when it was compared with that without nitrogen source. When mixed culture of white and yellow colony forming cells were applied to the soil column system contaminated by diesel, 30 mL/min of air flow rate was found to be sufficient to degrade diesel oil. The diesel degradation did not increase noticeably at higher flow rate. The addition of nitrogen source resulted in the increase in diesel degradability.

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Construction of 3D Culture Medium with Elastin-like Polypeptide (ELP) Hydrogel for Human Pluripotent Stem Cells

  • Lee, Jonghwan;Rhee, Ki-Jong;Jung, Donjgu
    • Biomedical Science Letters
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    • v.19 no.1
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    • pp.41-47
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    • 2013
  • Pluripotent stem cells (PSCs) have lots of potential in biomedical sciences owing to its potential to differentiate into any kind of cells in the body. However, it is still a challenge to culture PSCs on a large scale for application to regenerative medicine. Herein, we introduce a synthetic polymer that enables large-scale suspension culture of human PSCs. By employing suspension culture, it became unnecessary to use conventional substrata such as mouse embryonic fibroblast (MEF) or Matrigel$^{TM}$, which are believed to be main causative sources of xenogeneic contamination in cultured human PSCs in vitro. Human PSCs were cultured in the medium in which elastin-like polypeptide (ELP) dissolved. The ELP in the medium became harden as temperature increases by transforming the medium into a semi-solid gel that supported growth of human PSCs in suspension. Gel-sol transition temperature of ELP can be adjusted by modifying the peptide sequence in which 5 amino acids, Val-Pro-Gly-Xaa-Gly, repeated sequentially. We constructed 3D suspension media having transition temperature around $33{\sim}35^{\circ}C$ using an ELP consisted of 40, 60, or 80 repeats of a monomer, which was Val-Pro-Gly-Val-Gly. Among the ELPs, ELP80 was chosen as the best ELP to support growth of human PSCs in suspension culture. This result suggests that the ELP80 can be a medium component for culturing human PSCs in large-scale.

North American ginseng influences adipocyte-macrophage crosstalk regulation of inflammatory gene expression

  • Garbett, Jaime;Wilson, Sarah A.F.;Ralston, Jessica C.;Boer, Anna A. De;Lui, Ed M.K.;Wright, David C.;Mutch, David M.
    • Journal of Ginseng Research
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    • v.40 no.2
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    • pp.141-150
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    • 2016
  • Background: Adipocyte-macrophage communication plays a critical role regulating white adipose tissue (WAT) inflammatory gene expression. Because WAT inflammation contributes to the development of metabolic diseases, there is significant interest in understanding how exogenous compounds regulate the adipocyte-macrophage crosstalk. An aqueous (AQ) extract of North American (NA) ginseng (Panax quinquefolius) was previously shown to have strong inflammo-regulatory properties in adipocytes. This study examined whether different ginseng extracts influence adipocyte-macrophage crosstalk, as well as WAT inflammatory gene expression. Methods: The effects of AQ and ethanol (EtOH) ginseng extracts ($5{\mu}g/mL$) on adipocyte and macrophage inflammatory gene expression were studied in 3T3-L1 and RAW264.7 cells, respectively, using real-time reverse transcription polymerase chain reaction. Adipose tissue organ culture was also used to examine the effects of ginseng extracts on epididymal WAT (EWAT) and inguinal subcutaneous WAT (SWAT) inflammatory gene expression. Results: The AQ extract caused significant increases in the expression of common inflammatory genes (e.g., Mcp1, Ccl5, Tnf-${\alpha}$, Nos2) in both cell types. Culturing adipocytes in media from macrophages treated with the AQ extract, and vice versa, also induced inflammatory gene expression. Adipocyte Ppar-${\gamma}$ expression was reduced with the AQ extract. The AQ extract strongly induced inflammatory gene expression in EWAT, but not in SWAT. The EtOH extract had no effect on inflammatory gene expression in either both cell types or WAT. Conclusion: These findings provide important new insights into the inflammo-regulatory role of NA ginseng in WAT.

Effects of Date and Growth Regulators on the Culture of' Immature Zygotic Embryos of North American Ginseng

  • Hovius, Marilyn H. Y.;Saxena, Praveen K.;Proctor, John T. A.
    • Journal of Ginseng Research
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    • v.31 no.1
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    • pp.14-22
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    • 2007
  • As the zygotic embryo of North American ginseng (Panax quinquefolius L.) matured during stratification over 203 days it grew from 0.75 to 5.2 mm. Embryo excision and culturing on media containing different concentrations of two growth regulators, gibberellic acid ($GA_3$, 1 to 10 ${\mu}M$) and benzyladenine (BA, 1 to 5 ${\mu}M$), during stratification, showed that shoot and root number and the shoot, root and cotyledon length increased with increased stratification time. Gibberellic acid was the more effective growth regulator for increasing shoot and root number and shoot, root and cotyledon lengths. Immature embryos (stratified for up to 63 days) needed growth regulators for further development. Cultures on $GA_3$ at the last culture date (stratified for 203 days) when embryos were mature, produced multiple shoots but there was no effect of $GA_3$ concentration. Benzyladenine inhibited shoot and root growth regardless of embryo stratification. Growth regulators had little effect on cotyledon length of mature embryos. Embryos cultured on $GA_3$ combined with BA were green on all culture dates whereas greening in the control and BA treatments increased with culture date. The BA treatments induced 100% swelling of the embryos on the final culture date while in the control and $GA_3$ treatments there was no swelling. There was little or no curling in the control and BA treatments and a linear decrease in curling with culture date in the $GA_3$ and $GA_3$ + BA treatments.

Isolation and Evaluation of Protective Effect against Fusarium Wilt of Sesame Plants of Antibiotic Substance from Bacillus polymyxa KB-8

  • Hyun, Jae-Wook;Kim, Young-Hoon;Lee, Yong-Se;Park, Won-Mok
    • The Plant Pathology Journal
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    • v.15 no.3
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    • pp.152-157
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    • 1999
  • An antibiotic compound was isolated from the culture of an antagonist against Fusarium oxysporum f. sp. sesami, Bacillus polymyxa strain KB-8, and tested for the control of Fusarium wilt of sesame in greenhouse conditions. Optimum conditions for culturing the antagonist to obtain the maximum antibiotic activity were determined using different culture media, initial medium acidity, and incubation periods for which yeast -malt extract agar with the initial acidity of pH 5 and over 13 days culture were best. Antibiotic substances extracted by methanol had 2 main fractions, KB-8A and KB-8B, in thin layer chromatography (OLC) with Rf values of 0.35 and 0.67 in a solvent system of chloroform : methanol = 7 : 3. The fraction KB-8A wa purified further by XAD-2, silica gel and Sephadex LH-20 column chromatography, and crystalization. Its minimum inhibitory concentrations (MICs) were $12.8\mu\textrm{g}$/ml for F. oxysporum and Alternaria mali, $6.4\mu\textrm{g}$/ml for Colletotrichum gloeosporioides and Rhizoctonia solani, and $3.2\mu\textrm{g}$/ml for Phytophthora capsici. Soil drenching of antibiotic KB-8A in the concentrations of $13.0\mu\textrm{g}$/ml and $26.0\mu\textrm{g}$/ml effectively inhibited the Fusarium wilt of sesame in a greenhouse test, which appeared to be comparable to the fungicide benlate of $6.5\mu\textrm{g}$ a. i./ml.

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New Isolation Technique and Culture System for Clinical Applications of Human Amniotic Epithelial Stem Cells (인간태반양막유래 상피줄기세포의 임상적용을 위한 새로운 세포분리 및 배양 기술)

  • Woo, Sang-Kyu;Jo, Jung-Yoon;Shin, Il-Seob;Kang, Sung-Keun;Ra, Jeong-Chan
    • Development and Reproduction
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    • v.13 no.4
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    • pp.271-280
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    • 2009
  • Human placenta is abundant source of adult stem cells. Especially, amniotic epithelial cells have stem cell characteristics, expressing surface markers normally present on embryonic stem cells and germ cells. However, culturing and expanding amniotic epithelial cells in vitro without feeder cells are difficult due to endogenous characteristics of epithelial cells. In the present study, amniotic epithelial cells are isolated and proliferated in several passages by applying dithiothreitol and a Rho-associated kinase inhibitor in culture media. The cultured amniotic epithelial cells showed the epithelial and stem cell characteristics. In conclusion, human placenta-derived amniotic epithelial stem cells can be a major source of stem cells for medical treatment of various diseases without any controversial issues.

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Cultural characteristics of fungal species associated with deterioration or foxing of paper and chemical removal (종이변색균류의 배양적 특성 및 화학적 방법에 의한 변색제거)

  • Jo, Seong-Eun;Kim, Yong-Tae;Jeong, So-Yeong;Jo, Byeong-Muk;Lee, Jong-Gyu
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2009.04a
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    • pp.295-303
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    • 2009
  • The annals of Joseon dynasty, especially the volumes of King SeJong(1418-1450 A.D.), were heavily deteriorated by fungi. Investigations on the deteriorating and foxing fungi were carried out. Fungal structures on the beeswax, which were coated on the both side of Han-Ji, were suspected to be involved in the deterioration, and were observed by SEM. Isolation and culturing of these fungi were tried by scrubing swab samples and placing on the artificial media. Culture-independent approaches were used to identify the fungal strains associated with damages of beeswax and foxing of the paper by the analyses based on DNA sequences data from the specific ITS region of rDNA regions. In addition, well-known paper staining fungi(PSF), i.e., Aspergillus terreus var. terreus, Fusarium oxysporum, Chaetomium globosum, Cladosporium cladosporioides, and Alternaria solani, were compared in the mycelial growth and stain on beeswax and papers under different environmental conditions (temperature, light, moisture, etc). Fungal strains isolated from the air samples in the storage room and shelves were identified as Irpex sp., Arthrinium sacchari, Cladosporium tenuissimum, Aspergillus sclerotiorum, Sistotrema brinkmannii, and Hypoxylon bovei var. microsporum The isolated strains were compared in growth and stain patterns on beeswax and papers(Han-Ji, Hwa-Ji, and Yang-Ji) whether these can cause damage or foxing on the annals or not.

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Development of Biofugicide for Control of Gray Mole Rot of Eggplant Caused by Botrytis cinerea, and Bioassay in the Greenhouse Condition (가지 잿빛공팜이병 방제용 생물농약 개발 및 방제효과)

  • 김철승;이재필;송주희;임은경;정순재;하상영;문병주
    • Journal of Life Science
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    • v.11 no.3
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    • pp.235-241
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    • 2001
  • To select the sntagonistic bacteria against B. cinerea, isolates were screened from the eggplant leaves and rhizosphere soils in the eggplnat fields in the greenhouse. W1 and P99 isolates were selected by the inhibition of mycelial growth of B. cinerea E12 in vitro test. These isolates, W1 and P99, were identified as Bacillus subtilis and Pseudomonas putida, respectively, by the Bergeys manual and API systems, For the formulation of the antagonistic bacteria, the media for the mass production were prepared with biji(soybean curd residues) or soybean flour. B. subtilis W1 or P. putida P99 was mass cultured in biji broth or soybean flour extrect broth and then soybean flour, corn starch flour, rice glutinous flour and biji flour as high molecular substrates were added. These mixtures were dried, grinded and formulated as brofungicides of wettable powder type. The assess the control effect of biofungicides against the infection of B. cinerea, six types of formulations were assayed at the pot culturing with eggplant in the greenhouse. According to the results, there were no significant differences among the formulation methods. However, P99S or PppB formulated with P. putida P99 showed the highest control values as 90.4% and 96.1%, respectively. Then. BSB or BSD formulated whit B. subtilis W1 were 80.8% and 83.0%, respectively. There afforementioned values were more effective than that of chemical fungicide. Ipro W.P which showed as 72.6%.

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