• Title/Summary/Keyword: culturing media

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Influence of co-culturing muscle satellite cells with preadipocytes on the differentiation of adipocytes and muscle cells isolated from Korean native cattle

  • Choi, Chang Weon
    • Korean Journal of Agricultural Science
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    • v.45 no.4
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    • pp.715-723
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    • 2018
  • The present study was done to investigate the effect of co-culturing muscle satellite cells (MSCs) and intramuscular preadipocytes (IPs) on the differentiation of adipocytes and muscle cells isolated from Korean native cattle. MSCs and IPs were single-cultured in 10% fetal bovine serum/Dulbecco's modified Eagles medium (FBS/DMEM) for 48 h followed by culturing in 5% FBS/DMEM as the growth media. Then, the growth media was replaced by differentiation media composed of 2% FBS/DMEM without any additives for the single- or co-culture of muscle cells and intramuscular adipocytes to induce the differentiation of both cell types. Cell differentiation was measured by morphological investigation and cytosolic enzyme analysis of glycerol-3-phosphate dehydrogenase (GPDH) for the adipocytes and creatine kinase (CK) for the muscle cells. In the morphological test, the presence of muscle cells did not stimulate adipocyte differentiation showing more differentiation of the adipocytes in the single-culture compared to the co-culture condition. However, the differentiation of muscle cells was promoted by adipocytes in the co-culture. The results of the enzymatic analysis were highly associated with the morphological results with a statistically higher GPDH activity (p < 0.05) appearing in the single-culture than in the co-culture, whereas the opposite was true for the CK activity of the muscle cells (p < 0.05). By manipulating in vivo the milieu using a co-culture, we could detect the difference in the rate of cell differentiation and suggest that a co-culture system is a more reliable and precise technique compared to a single-culture. Further studies on various co-culture trials including supplementation of differentiating substances, gene expression analysis, etc. should be done to obtain practical and fundamental data.

Growth Rate and Antimicrobial Activity of Lactobacillus plantarum Reacting with Bacterial Culture Supernatant (세균 배양액과 반응한 Lactobacillus plantarum의 생장속도 및 항균 활성에 대한 연구)

  • Ha, Eun-Mi
    • Microbiology and Biotechnology Letters
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    • v.43 no.3
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    • pp.187-194
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    • 2015
  • In this study, L. plantarum, when reacting with the culture media of potential pathogenic bacteria, exhibited an increase in growth rate and antimicrobial activity. In order to examine the characteristics and the nature of the reaction with the bacteria, this study carried out experiments involving culturing the test bacteria in M9 minimal media. Subsequently, the supernatant was incrassated by the decompression-drying method. Through colony forming unit assay, it was confirmed that L. plantarum had the function of growth inhibition to various bacteria. After culturing L. plantarum with bacterial media, the growth rate of L. plantarum was measured by absorbance (OD600), the results showed that the growth rate (E. coli treatment group: OD600 = 0.848, S. typhimurium treatment group: OD600 = 0.848) increased, as compared with the non-treated control group (OD600 = 0.48). In contrast, the concentrate itself did not induce the growth of L. plantarum. These results were observed as a universal phenomenon of the Lactobacillus species. Moreover, the increase in antimicrobial activity was observed in L. plantarum, which reacted with the culture media of E. coli and S. typhimurium, through a disc diffusion assay, and the result of growth inhibition against various bacteria was induced. Finally, based on the analysis results of the characteristics of bacteria culture media, which increased the growth rate of L. plantarum and antibacterial activity, the bacterial media had a tolerance for catabolic enzymes, pH 2−8 and heat. Therefore, this substance can be said to be a small molecule which is highly stable under various conditions.

Transcriptional Interplay between Malassezia restricta and Staphylococcus Species Co-Existing in the Skin Environment

  • Hyun Oh Yang;Yong-Joon Cho;Jae Min Lee;Kyoung-Dong Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.3
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    • pp.319-328
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    • 2023
  • Malassezia and Staphylococcus are the most dominant genera in human skin microbiome. To explore the inter-kingdom interactions between the two genera, we examined the transcriptional changes in Malassezia and Staphylococcus species induced upon co-culturing. RNA-seq analyses revealed that genes encoding ribosomal proteins were upregulated, while those encoding aspartyl proteases were downregulated in M. restricta after co-culturing with Staphylococcus species. We identified MRET_3770 as a major secretory aspartyl protease coding gene in M. restricta through pepstatin-A affinity chromatography followed by mass spectrometry and found that the expression of MRET_3770 was significantly repressed upon co-culturing with Staphylococcus species or by incubation in media with reduced pH. Moreover, biofilm formation by Staphylococcus aureus was inhibited in the spent medium of M. restricta, suggesting that biomolecules secreted by M. restricta such as secretory aspartyl proteases may degrade the biofilm structure. We also examined the transcriptional changes in S. aureus co-cultured with M. restricta and found co-cultured S. aureus showed increased expression of genes encoding ribosomal proteins and downregulation of those involved in riboflavin metabolism. These transcriptome data of co-cultured fungal and bacterial species demonstrate a dynamic interplay between the two co-existing genera.

Optimal Culturing and Enhancement of Lipid Accumulation in a Microalga Botryococcus braunii (미세조류 Botryococcus braunii의 배양조건 최적화 및 지질축적 향상)

  • Kwon, Sung-Hyun;Lee, Eun-Mi;Cho, Dae-Chul
    • Journal of Environmental Science International
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    • v.21 no.7
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    • pp.779-785
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    • 2012
  • Several tests and experimental work have been done for identifying the best growth conditions and accumulated amount of lipid moiety in B. braunii, a microalga(UTEX 572) in terms of media composition. The specific growth rate was found to be the highest at 0.15 g/L-day when the phosphorus concentration was doubled with the other ingredients at the normal level. Experiments for varied media compositions revealed that the accumulation of lipid was the highest at 48% (dry cell weight based) in the nitrogen deficient medium and its corresponding specific growth rate was comparative to that in the normal BG 11 medium. In the bubble column experiments, carbon dioxide containing air produced four times more cell mass than air only. Light and glucose addition also enhanced cell mass with maximum, 1.8 g/L and accordingly 42% of lipid composition, which turned out to be a better strategy for higher lipid-producing microalgal culture.

In vitro propagation of Phaleonopsis hybrid 'Little gem' by culturing apical part and axillary bud of flower stalk

  • Chung, Mi Young;Naing, Aung Htay;Khatun, Khadiza;Ahn, Hyung Geun;Lim, Ki Byung;Kim, Chang Kil
    • Journal of Plant Biotechnology
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    • v.43 no.4
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    • pp.438-443
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    • 2016
  • The in vitro propagation of the commercially important Phalaeonopsis hybrid 'Little gem' was achieved by culturing the apical part and axillary buds excised from flower stalks. The explants were cultured on 5 different basal media: $3.0{\cdot}L^{-1}$ Hyponex and $4.0{\cdot}L^{-1}$ peptone ($H_3P_4$) and Murashige & Skoog (MS) media were shown to be suitable for shoot regeneration. The MS medium supplemented with $5.0mg{\cdot}L^{-1}$ 6-benzylaminopurine (BA) was found to be more efficient for shoot regeneration. However, the number of shoots induced by axillary buds was higher than that induced by the apical part. Incubation of the apical part under darkness for one week, as well as of the explants in the same medium with activated charcoal (AC) $0.5g{\cdot}L^{-1}$ promoted shoot regeneration and shoot growth; similar growth was not observed with axillary buds.

Diagnosis of Tuberculosis; Serodiagnosis and Molecular Biologic Approach (결핵진단의 면역학적 및 분자생물학적 방법)

  • Shin, Wan-Shik
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.1
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    • pp.1-6
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    • 1992
  • The diagnosis of tuberculosis is usually established using staining and culturing techniques. Fluorescent stains have improved the sensitivity of direct microscopy. Improved culture media coupled with radiometric means of detecting early mycobacterial growth have shortened the time needed for cultural diagnosis. Rapid immunodiagnostic techniques based on the detection of mycobacterial antigen or of antibodies to theses antigens have not, however, come into widespread clinical use. The DNA or RNA hybridization tests with labeled specific probes which have been described so far are not sensitive enough to be used for clinical speicimens without prior culturing. The advent of the polymerase chain reaction (PCR) has opened new possibilities for diagnosis of microbial infections. This technique has already been applied to a number of microorganisms. In the field of mycobacteria the PCR has been used to identify and to detect DNAs extracted from various mycobacteria. However, despite the extraordinary enthusiasm surrounding this technique and the considerable investiment, PCR has not emerged from the developmental "trenches" in the passed several years. It may be a considerable lenth of time before clinical microbiology laboratories become PCR playgrounds because many details remain to be worked out.

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Production of Taxane Derivatives by Cell Culture of Korean Taxus Species (I) (한국산 주목의 세포배양에 의한 Taxane 유도체의 생산 (I))

  • Kim, You-Sun;Shin, Seung-Won
    • Korean Journal of Pharmacognosy
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    • v.27 no.3
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    • pp.262-266
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    • 1996
  • Undifferentiated callus were induced from the young leaves of Taxus cuspidata by treatment with various combinations of plant growth hormones. The effects of light and culturing temperature on production of baccatin III, the precursor of taxol, were studied. The contents of baccatin III in the cultured callus were analysed by HPLC. The illumination of fluorescence and administration of isoleucine, one of the possible substrates in biosynthesis of terpenoids, to the culturing media increased the production of baccetin III.

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Studies on Culture of Leptospires On Growth of Leptospires in Cox's Liquid Media and Korthof's Media, Added Rabbit Serum and Thiamine Hydrochloride, Inactivated at High Temperature (Leptospira 균(菌) 배양(培養)에 관한 연구(硏究) -가열처리(加熱處理)한 토끼 혈청가배지(血淸加培地)와 Thiamine 첨가배지(添加培地)에서의 Leptospira 균(菌)의 발육(發育)에 관하여-)

  • Seo, Ik Soo
    • Korean Journal of Veterinary Research
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    • v.15 no.1
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    • pp.75-81
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    • 1975
  • The rate of growth of Leptospires were studied by culturing in Korthof's media and Cox's liquid media which were treated at four different temperatures. The effect of thiamine hydrochloride in the culture of leptospires were also studied. The results obtained were summerized as followings: 1. The best growth of Leptospira icterohaemorrhagiae and Leptospira canicola was observed in Korthof's media with the rabbit serum heated for 60 minutes at $56^{\circ}C$. 2. A good degree of growth of Leptospira icterohaemorrhagiae and Leptospira canicola was observed in both Cox's liquid media with rabbit serum heated for 30 minutes at $100^{\circ}C$ and with fresh rabbit serum. 3. The growth of Leptospira was enhanced by adding small amount of thiamine to the both media which were shown to be unsatisfactory for the growth of the organisms. 4. Culture of leptospires were attained by simply heating for 30 minutes at $100^{\circ}C$ both Korthof's media and Cox's liquid media containing rabbit serum obtained without aspetic procedures.

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Root and Shoot Formation in Explant and Callus Derived from Root and Cotyledon of GinBeng(Panun ginseng C. A. Meyer) (인삼근 및 자엽 Callus의 기관분화에 관한 연구)

  • Choe, Gwang-Tae;Kim, Myeong-Won;Sin, Hui-Seok
    • Journal of Ginseng Research
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    • v.5 no.1
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    • pp.35-40
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    • 1981
  • Explants of mature root tissues and calli derived from root and cotyledon of Panax ginseng were cultured in vitro on Murashige and Skoog medium supplemented with 2, 4-dichlorophen-oxyacetic acid(3,4-D), naphthaleneacetic acid(NAA), benzyladenine, and gibberellic acid to assess their capacity to regenerate organs. Root formation at high percentage (46.2-61.1%) was obtained 20-30 days after culturing on media supplemented with combinations of NAA(5 mg/l) and kinetin (1 mg/l), And calli derived from cotyledon produced numerous embryoids in media($\frac{1}{2}$MS) containing 2,4-D(0.5 mg/l) and kinetin (0.5 mg/l). Reculture of these embryoids in media($\frac{1}{2}$MS) enriched with 1 mg/l of benzyladenine and 1 mg/l of gibberellic acid resulted in more plantlet regeneration.

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