• Title/Summary/Keyword: Optimal culture conditions

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Surface maker and gene expression of human adipose stromal cells growing under human serum. (인체혈청 하에서 배양한 인체지방기질줄기세포의 표면항원 및 유전자 발현)

  • Jun, Eun-Sook;Cho, Hyun-Hwa;Joo, Hye-Joon;Kim, Hoe-Kyu;Bae, Yong-Chan;Jung, Jin-Sup
    • Journal of Life Science
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    • v.17 no.5 s.85
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    • pp.678-686
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    • 2007
  • Human mesenchymal stem cells(hMSC), that have been reported to be present in bone marrow, adipose tissues, dermis, muscles and peripheral blood, have the potential to differentiate along different lineages including those forming bone, cartilage, fat, muscle and neuron. Therefore, hMSC are attractive candidates for cell and gene therapy. The optimal conditions for hMSC expansion require medium supplemented with fetal bovine serum(FBS). Some forms of cell therapy will involve multiple doses, raising a concern over immunological reactions caused by medium-derived FBS proteins. Previously, we have shown that hADSC can be cultured in human serum(HS) during their isolation and expansion, and that they maintain their proliferative capacity and ability for multilineage differentiation and promote engraftment of peripheral blood-derived CD34 cells mobilized from bone marrow in NOD/SCID mice. In this study we determined whether hADSC grown in HS maintain surface markers expression similar with cells grown in FBS during culture expansion and compared gene expression profile by Affymetrix microarray. Flow cytometry analysis showed that HLA-DR, CD117, CD29 and CD44 expression in HS-cultured hADSC during culture expansion were similar with that in FBS-cultured cells. However, the gene expression profile in HS-cultured hADSC was significantly different from that in FBS-cultured cells. Therefore, these data indicated that HS-cultured hADSC should be used in vivo animal study of hADSC transplantation for direct extrapolation of preclinical data into clinical application.

Isolation and Characterization of a Marine Bacterium, Pseudomonas sp. YJ-1 with Anti-Methicillin Resistant Staphylococcus aureus Activity (항 Methicillin Resistant Staphylococcus aureus 활성을 가지는 해양미생물 Pseudomonas sp. YJ-1의 분리와 특성)

  • Woo, Ye-Ju;Jeong, Seong-Yun
    • Korean Journal of Environmental Biology
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    • v.35 no.4
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    • pp.694-705
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    • 2017
  • The aim of this study was to isolate and identify marine bacterium with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity, and to purify the anti-MRSA compound, as well as to determine its activity and synergistic effects. Among the marine bacteria isolated in this study, the YJ-1 isolate had the strongest anti-MRSA activity. The YJ-1 isolate was identified on the basis of its biochemical characteristics and an analysis of 16S rRNA gene sequences. The YJ-1 isolate showed over 99.2% homology with Pseudomonas stutzeri, and was designated as a Pseudomonas sp. YJ-1. The optimal culture conditions were $25^{\circ}C$ and initial pH 7.0. For the purification of the anti-MRSA compounds, the YJ-1 was cultured in Pa PES-II medium, and the culture filtrates were extracted by ethyl acetate, hexane, and 80% MeOH. The 80% MeOH fraction was separated by a $C_{18}$ ODS column, silica gel chromatography and a reverse phase HPLC, to yield three anti-MRSA agents, the MR1, MR2, and MR3 compounds. When the MR1 compound of $250{\mu}g\;mL^{-1}$ concentration was applied to the MRSA cells, over 95% of bacterial cells was killed within 48 hr. Compared with vancomycin and ampicillin, the MR1 compound showed significant anti-MRSA activity. In addition, the anti-MRSA activity was increased by dose and time dependent manners. Furthermore, the combination of an MR1 compound with vancomycin produced a more rapid decrease in the MRSA cells than did the MR1 compound alone. Taken together, our results suggest that the Pseudomonas sp. YJ-1 and its anti-MRSA compounds could be employed as a natural antibacterial agent in MRSA infections.

Optimizing In Vitro Propagation of Sophora koreensis Nakai using Statistical Analysis (다양한 통계분석 기법을 이용한 개느삼(Sophora koreensis Nakai)의 기내 증식 최적 조건 구명)

  • Jeong, Ukhan;Lee, Hwa;Park, Sanghee;Cheong, Eun Ju
    • Journal of Korean Society of Forest Science
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    • v.110 no.1
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    • pp.53-63
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    • 2021
  • Sophora koreensis Nakai is an indigenous plant in Koreawith a restricted natural range, part of which is in Gangwon province. The species is known to contain phytochemicals that have beneficial effects on human health, and it is economically important in bioindustry. Because of the limited number of plants in a small range of habitats, the mass-propagation method should be developed for use and conservation. In vitro tissue culture is a reliable method in terms of mass propagation from selected clones of the species. We investigated the optimal conditions of the medium in this process, especially focusing on the concentrations of plant growth regulators(PGRs) in the culture of stem-containing axillary buds. Three statistical methods, i.e., ANOVA, response surface method(RSM), and fuzzy clustering were used to analyze the plant growth, number of shoots induced, and shoot length with various combinations of PGRs. Results from the RSM differed from those of the other two methods; thus, the method was not suitable. ANOVA and fuzzy clustering showed similar results. However, more accurate results were obtained using fuzzy clustering because it provided a probability for each treatment. On the basis of the fuzzy clustering analysis, stem tissue produced the greatest number of shoots(11.03 per explant; 63.33%) on a medium supplemented with 5-��M 6-benzylaminopurine and 2.5-��M thidiazuron(TDZ). Proliferation of shoots(2.18 ± 0.21 cm, 63.33%) was attained on a medium supplemented with 2.5-��M BA, 2.5-��M TDZ, and 2.5-��M gibberellic acid.

Development of an Efficient In Vitro Screening Method for Selection of Resistant Lily Cultivars Against Fusarium oxysporum f. sp. lilii (백합 구근부패병 저항성 검정을 위한 기내 간편 검정법 개발)

  • Jang, Ji-Young;Moon, Ki-Beom;Ha, Jang-Ho;Park, Ji-Sun;Kim, Mi-Jin;Jeon, Jae-Heung;Lee, Geung-Joo;Kim, Hyun-Soon
    • Horticultural Science & Technology
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    • v.33 no.6
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    • pp.883-890
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    • 2015
  • The soilborne fungus Fusarium oxysporum f. sp. lilii (Fol) is a serious threat to all lily cultivars, especially infecting bulbs and flowers. It has become increasingly important to develop varieties resistant against the bulb rot disease. Genetic diversity of cultivars and reliable screening methods are required for this purpose. Here, an efficient in vitro screening system for evaluating resistance to Fol in 38 in vitro-grown lily plants was investigated. Various factors including culture conditions of Fol, inoculum density, appropriate plant materials, inoculation method and duration, and incubation period of plant materials after inoculation were combined to optimize the screening method. As a result, we suggest optimal conditions for an in vitro screening system for the selection of Fol-resistant lily cultivars as follows. Fol was grown on potato dextrose agar (PDA) medium for 6 days at $25^{\circ}C$ in darkness and used as working inoculation. Spore suspensions were prepared (inoculum density: $1.0{\times}10^4$ $spores{\cdot}mL^{-1}$), and then leaf segments $1.5{\times}2.0cm^2$ were inoculated by dipping for 22 hours at $25^{\circ}C$ in dark. Later, leaves were cultured on 0.6% agar plates at $25^{\circ}C$ and 50% humidity with a photoperiod of 16 hours light/8 hours dark (fluence rate of $40{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) to examine the progress of bulb rot. After 7 days, disease levels were classified into indices 1 (no symptom) to 6 (serious bulb rot). Soil inoculation of Fol carried out with resistant or susceptible lily cultivars that had been selected through in vitro screening confirmed the reproducibility of results. Therefore, the in vitro screening method established in this study is efficient and reliable for selection of lily cultivars resistant against bulb rot disease.

A study on the CRM strategy for medium and small industry of distribution (중소유통업체의 CRM 도입방안에 관한 연구)

  • Kim, Gi-Pyoung
    • Journal of Distribution Science
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    • v.8 no.3
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    • pp.37-47
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    • 2010
  • CRM refers to the operating activities that always maintain and promote good relationship with customers to ultimately maximize the company's profits by understanding the value of customers to meet their demands, establishing a strategy which may maximize the Life Time Value and successfully operating the business by integrating the customer management processes. In our country, many big businesses are introducing CRM initiatively to use it in marketing strategy however, most medium and small sized companies do not understand CRM clearly or they feel difficult to introduce it due to huge investment needed. This study is intended to present CRM promotion strategy and activities plan fit for the medium and small sized companies by analyzing the success factors of the leading companies those have already executed CRM by surveying the precedents to make the distributors out of the industries have close relation with consumers to overcome their weakness in scale and strengthen their competitiveness in such a rapidly changing and fiercely competing market. There are 5 stages to build CRM such as the recognition of the needs of CRM establishment, the establishment of CRM integrated database, the establishment of customer analysis and marketing strategy through data mining, the practical use of customer analysis through data mining and the implementation of response analysis and close loop process. Through the case study of leading companies, CRM is needed in types of businesses where the companies constantly contact their customers. To meet their needs, they assertively analyze their customer information. Through this, they develop their own CRM programs personalized for their customers to provide high quality service products. For customers helping them make profits, the VIP marketing strategy is conducted to keep the customers from breaking their relationships with the companies. Through continuous management, CRM should be executed. In other words, through customer segmentation, the profitability for the customers should be maximized. The maximization of the profitability for the customers is the key to CRM. These are the success factors of the CRM of the distributors in Korea. Firstly, the top management's will power for CS management is needed. Secondly, the culture across the company should be made to respect the customers. Thirdly, specialized customer management and CRM workers should be trained. Fourthly, CRM behaviors should be developed for the whole staff members. Fifthly, CRM should be carried out through systematic cooperation between related departments. To make use of the case study for CRM, the company should understand the customer and establish customer management programs to set the optimal CRM strategy and continuously pursue it according to a long-term plan. For this, according to collected information and customer data, customers should be segmented and the responsive customer system should be designed according to the differentiated strategy according to the class of the customers. In terms of the future CRM, integrated CRM is essential where the customer information gathers together in one place. As the degree of customers' expectation increases a lot, the effective way to meet the customers' expectation should be pursued. As the IT technology improved rapidly, RFID (Radio Frequency Identification) appears. On a real-time basis, information about products and customers is obtained massively in a very short time. A strategy for successful CRM promotion should be improving the organizations in charge of contacting customers, re-planning the customer management processes and establishing the integrated system with the marketing strategy to keep good relation with the customers according to a long-term plan and a proper method suitable to the market conditions and run a company-wide program. In addition, a CRM program should be continuously improved and complemented to meet the company's characteristics. Especially, a strategy for successful CRM for the medium and small sized distributors should be as follows. First, they should change their existing recognition in CRM and keep in-depth care for the customers. Second, they should benchmark the techniques of CRM from the leading companies and find out success points to use. Third, they should seek some methods best suited for their particular conditions by achieving the ideas combining their own strong points with marketing. Fourth, a CRM model should be developed that will promote relationship with individual customers just like the precedents of small sized businesses in Switzerland through small but noticeable events.

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Mycelial Culture and Fruiting Analysis of Panellus edulis Strains Collected in Korea (Panellus edulis 수집 균주의 균사배양 및 자실체 특성 분석)

  • Woo, Sung-I;Ryoo, Rhim;Jang, Yeongseon;Park, Youngae;Jeong, Yeun Sug;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.46 no.3
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    • pp.281-294
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    • 2018
  • Molecular analysis using the internal transcribed spacer region sequences revealed that the strains used in this study, which were formerly identified as Panellus serotinus, are Panellus edullis. After Universal Fungal PCR Fingerprinting (UFPF) analysis, eight strains of P. edulis were divided into two groups. We conducted fundamental research on mycelial growth and sawdust cultivation to understand the cultural characteristics of eight wild P. edulis strains collected from Korean forests. All strains showed faster and denser mycelial growth on potato dextrose agar (PDA) than on other media (malt extract agar, Sabouraud dextrose agar). Optimal conditions for mycelial growth were: $20^{\circ}C$ on PDA, $25^{\circ}C$ on potato dextrose broth (PDB), and pH 5~8 on PDB at $25^{\circ}C$. Two strains (NIFoS 2407, 3993) were selected as excellent strains based on mycelial growth and density on PDA. NIFoS 2792 showed high cellulase activities on carboxymethyl cellulose (CMC) agar, and NIFoS 2387 and 2804 exhibited high laccase activities on ABTS-containing agar media. The mycelial growth of P. edulis was the fastest on Quercus acutissima and Q. mongolica sawdust media, and mycelial density was the highest on Quercus spp. sawdust-containing media. Sawdust cultivation of P. edulis was successful. The conditions were 80~85 days of cultivation period after spawn inoculation, 10~11 days for primordial formation at $17{\sim}18^{\circ}C$, and 15~20 days for fruiting growth. NIFoS 2804 and 3993 were selected as good strains in terms of cultivation period and mushroom production. These results could be useful for the artificial cultivation of P. edulis.

Growth responses of kelp species Ecklonia cava to different temperatures and nitrogen sources (온도와 질소원 종류에 따른 대형갈조류 감태(Ecklonia cava)의 생장)

  • Choi, Sun Kyeong;Kang, Yun Hee;Park, Sang Rul
    • Korean Journal of Environmental Biology
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    • v.38 no.3
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    • pp.404-415
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    • 2020
  • We investigated the seasonal variations in growth and physiological responses of the kelp species Ecklonia cava to different nitrogen sources to establish indoor culture conditions for mass production. Ecklonia cava was cultivated for 10 days in 16 combinations of seawater temperatures (15, 17, 21, and 25℃) and different nitrogen sources (control; NH-NH4+ 100 μM; NO-NO3- 100 μM; and NHNO-NH4+ 50 μM+NO3- 50 μM). The growth and growth rate of the blade were affected by temperature. The mean fresh weight and area-based daily growth rate were the highest (5.8±0.5 and 6.6±0.5% day-1, respectively) at 15℃ and the lowest (2.2±0.2 and 3.0±0.3% day-1, respectively) at 25℃. The daily growth rate was the highest in the NH and NO treatments and lowest in the control. The nitrate reductase activity of E. cava varied with water temperature (season). The highest activity was in the control (1.32±0.10 μmol NO2- g-1 dry weight h-1) and the lowest was in the NH treatment (0.25±0.02 μmol NO2- g-1 dry weight h-1). The photosynthetic pigment concentrations reached a maximum value in the NHNO treatment and a minimum value in the control. These results showed that water temperature played an important role in the cultivation of E. cava and that a single supply of NH4+ or NO3- may induce the accelerated growth of E. cava. The growth and physiological responses of E. cava to different nitrogen sources during each season provide valuable information for determining the optimal nitrogen source in E. cava cultivation under indoor conditions.

Biological Characteristics and Growth of the Korean Freshwater Rotifer, Brachionus calyciflorus at Various Temperatures (한국 담수산 윤층 Brachionus calyciflorus의 생물학적 특징과 온도별 성장)

  • 강언종;이배익;김응오
    • Journal of Aquaculture
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    • v.10 no.4
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    • pp.449-456
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    • 1997
  • This study was concuted to determine the optimal conditions for raising the freshwater rotifer, Brachinus calyciflorus. The authors presented some biological informatin obtained from incubation experiment under the various controlled temperatures. Lorica size of the rotifer was divided into two groups : the length and the width for the S-type was $141.0\pm16.7\mu m$($110.1-182.5\;\mu m, n=44$)and $107.0\pm20.3\mu m\;(75.3-152.3\mu m, n=44)$, and those for the L-type was $262.8\pm15.2\mu m\;(234.4-288.6\mu m,\;n=20)\;and\;182.6\pm13.4\mu m (159.8-207.0\mu m,\;n=20$), respectively. The number of eggs being attached on the female varied from 1 to 11 at various culture conditions. Egg type was divided into two groups, large and small. Large and small egg was measured in its major axis as 85a.7-107.8$\mu$m and 55.1-65.2$\mu$m for S-type, and 104.9-121.8 $\mu$m and 62.8-89.1$\mu$m for L-type respectively. The maximum density was reached at 4th day after incubation. The density was 583.9 rotifers/$m\ell$ for $25^{\circ}C$-experimental. group and 421.3 rotifers/$m\ell$ for $22^{\circ}C$-experimental. group respectively. In the case of $28^{\circ}C$-experimental. group, it suddenly decreased into 4.7 rotifers/$m\ell$ at 1st day after incubations and did not recover to its initial density. The maximum rate of increase of populatin per day was reached 0.802 for $22^{\circ}C$-experimental. group at day 2 and fluctuated thereafter. For $25^{\circ}C$-experimental. group it increased to 0.964 at day 3 of incubation and then declined. And the egg ratio of female was reached the maximum of 0.614 for 22$^{\circ}C$- at 3rd day and 0.772 for $25^{\circ}C$-experimental. group at 4th day of incubation.

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Purification of Human HtrA1 Expressed in E. coli and Characterization of Its Serine Protease Activity (E. coli에서 발현된 human HtrA1 단백질의 정제와 HtrA1의 serine protease 활성 조건에 관한 연구)

  • Kim, Kyung-Hee;Kim, Sang-Soo;Kim, Goo-Young;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1133-1140
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    • 2006
  • Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.

The Optimal Production and Characteristics of an Alginate-degrading Enzyme from Vibrio sp. PKA 1003 (Vibrio sp. PKA 1003의 알긴산 분해 조효소 생산 최적 조건과 조효소의 특성)

  • Kim, Hyun-Jee;Kim, Koth-Bong-Woo-Ri;Kim, Dong-Hyun;SunWoo, Chan;Jung, Seul-A;Jeong, Da-Hyun;Jung, Hee-Ye;Lim, Sung-Mee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.434-440
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    • 2013
  • This study was conducted to screen the characteristics and alginate-degrading activity of marine bacterium isolated from brown seaweed (Sargassum thunbergii). The results of 16S ribosomal RNA sequence analysis the strain the genus Vibrio and the strain was subsequently named Vibrio sp. PKA 1003. The optimum culture conditions for the growth of Vibrio sp. PKA 1003 were at pH 7, 3% NaCl, $25^{\circ}C$, and 6% alginic acid, with a 48-hour incubation time. A crude enzyme preparation from Vibrio sp. PKA 1003, showed its highest levels of alginate-degradation activity when cultured at pH 9, $30^{\circ}C$, and 6% alginic acid, with a 63-hour incubation time. Thin layer chromatography analyses confirmed that the crude enzyme released monomers or oligomers from sodium alginate, and results from trypsin treatment showed that the alginate degrading activity depends on this enzyme produced by Vibrio sp. PKA 1003. These results suggest that Vibrio sp. PKA 1003 and its alginate-degrading crude enzyme is useful for the production of alginate oligosaccharides.