• Title/Summary/Keyword: soft agar

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Tumorigenicity Evaluation of Umbilical Cord Blood-derived Mesenchymal Stem Cells

  • Park, Sang-Jin;Kim, Hyun-Jung;Kim, Woojin;Kim, Ok-Sun;Lee, Sunyeong;Han, Su-Yeon;Jeong, Eun Ju;Park, Hyun-shin;Kim, Hea-Won;Moon, Kyoung-Sik
    • Toxicological Research
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    • v.32 no.3
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    • pp.251-258
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    • 2016
  • Mesenchymal stem cells (MSCs) have been identified in multiple types of tissue and exhibit characteristic self-renewal and multi-lineage differentiation abilities. However, the possibility of oncogenic transformation after transplantation is concerning. In this study, we investigated the tumorigenic potential of umbilical cord blood-derived MSCs (hUCB-MSCs) relative to MRC-5 and HeLa cells (negative and positive controls, respectively) both in vitro and in vivo. To evaluate tumorigenicity in vitro, anchorage-independent growth was assessed using the soft agar colony formation assay. hUCB-MSCs and MRC-5 cells formed few colonies, while HeLa cells formed a greater number of larger colonies, indicating that hUCB-MSCs and MRC-5 cells do not have anchorage-independent proliferation potential. To detect tumorigenicity in vivo, hUCB-MSCs were implanted as a single subcutaneous injection into BALB/c-nu mice. No tumor formation was observed in mice transplanted with hUCB-MSCs or MRC-5 cells based on macro- and microscopic examinations; however, all mice transplanted with HeLa cells developed tumors that stained positive for a human gene according to immunohistochemical analysis. In conclusion, hUCB-MSCs do not exhibit tumorigenic potential based on in vitro and in vivo assays under our experimental conditions, providing further evidence of their safety for clinical applications.

Chemical Transformation of Human Keratinocytes by 2,3,7,8-Tetrachlorodibenxo-$\rho$-dioxin

  • Kang, Mi-Kyung;Choi, Young-Sill;Ryeom, Tai-Kyung;Eom, Mi-Ok;Park, Mi-Sun;Jee, Seung-Won;Kim, Kang-Ryune;Kim, Ok-Hee;Kang, Ho-Il
    • Environmental Mutagens and Carcinogens
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    • v.26 no.3
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    • pp.69-76
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    • 2006
  • 2,3,7,8-Tetrachlorodibenzo-$\rho$-dioxin(TCDD) is a ubiquitous, persistent environmental contaminant and the most powerful carcinogen categorized by IARC. Although the mechanism of carcinogenesis by TCDD is poorly understood, several studies have shown that the skin is one of target organs far TCDD. In this study, we investigated the neoplastic transformation of human keratinocyte-derived cell line, HaCaT, by chemical transformation method using N-methyl-N'-nitro-N-nitrorsoguanidine(MNNG) and TCDD. We found that subsequent exposure to TCDD for 3 weeks after initial exposure to MNNG markedly induced transformed cells. It was suggested that TCDD can act as a potent promoter in HaCaT cells. Furthermore, these transformed cells showed morphological alternations in soft agar and increased telomerase activity. Therefore, the TCDD treatment of HaCaT cells by initiated with MNNG could promote neoplastic transformation without stimulation by exogenous growth factors. As a result, TCDD had a strong potency as a promoter in nontumorigenic immortalized human epidermal keratinocytes.

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Inhibitory activities of Perilla frutescens britton leaf extract against the growth, migration, and adhesion of human cancer cells

  • Kwak, Youngeun;Ju, Jihyeung
    • Nutrition Research and Practice
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    • v.9 no.1
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    • pp.11-16
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    • 2015
  • BACKGROUND/OBJECTIVES: Perilla frutescens Britton leaves are a commonly consumed vegetable in different Asian countries including Korea. Cancer is a major cause of human death worldwide. The aim of the current study was to investigate the inhibitory effects of ethanol extract of perilla leaf (PLE) against important characteristics of cancer cells, including unrestricted growth, resisted apoptosis, and activated metastasis, using human cancer cells. MATERIALS/METHODS: Two human cancer cell lines were used in this study, HCT116 colorectal carcinoma cells and H1299 non-small cell lung carcinoma cells. Assays using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide were performed for measurement of cell growth. Soft agar and wound healing assays were performed to determine colony formation and cell migration, respectively. Nuclear staining and cell cycle analysis were performed for assessment of apoptosis. Fibronectin-coated plates were used to determine cell adhesion. RESULTS: Treatment of HCT116 and H1299 cells with PLE resulted in dose-dependent inhibition of growth by 52-92% (at the concentrations of 87.5, 175, and $350{\mu}g/ml$) and completely abolished the colony formation in soft agar (at the concentration of $350{\mu}g/ml$). Treatment with PLE at the $350{\mu}g/ml$ concentration resulted in change of the nucleus morphology and significantly increased sub-G1 cell population in both cells, indicating its apoptosis-inducing activity. PLE at the concentration range of 87.5 to $350{\mu}g/ml$ was also effective in inhibiting the migration of H1299 cells (by 52-58%) and adhesion of both HCT116 and H1299 cells (by 25-46%). CONCLUSIONS: These results indicate that PLE exerts anti-cancer activities against colon and lung cancers in vitro. Further studies are needed in order to determine whether similar effects are reproduced in vivo.

Growth, Clonability, and Radiation Resistance of Esophageal Carcinoma-derived Stem-like Cells

  • Li, Jian-Cheng;Liu, Di;Yang, Yan;Wang, Xiao-Ying;Pan, Ding-Long;Qiu, Zi-Dan;Su, Ying;Pan, Jian-Ji
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4891-4896
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    • 2013
  • Objective: To separate/enrich tumor stem-like cells from the human esophageal carcinoma cell line OE-19 by using serum-free suspension culture and to identify their biological characteristics and radiation resistance. Methods: OE-19 cells were cultivated using adherent and suspension culture methods. The tumor stem-like phenotype of CD44 expression was detected using flow cytometry. We examined growth characteristics, cloning capacity in soft agar, and radiation resistance of 2 groups of cells. Results: Suspended cells in serum-free medium formed spheres that were enriched for CD44 expression. CD44 was expressed in 62.5% of suspended cells, but only in 11.7% of adherent cells. The suspended cells had greater capacity for proliferation and colony formation in soft agar than the adherent cells. When the suspended and adherent cells were irradiated at 5 Gy, 10 Gy, or 15 Gy, the proportion of CD44+ suspended cells strongly and weakly positive for CD44 was 77.8%, 66.5%, 57.5%; and 21.7%, 31.6%, 41.4%, respectively. In contrast, the proportion of CD44+ adherent cells strongly positive for CD44 was 18.9%, 14.%, and 9.95%, respectively. When the irradiation dose was increased to 30 Gy, the survival of the suspended and adherent cells was significantly reduced, and viable CD44+ cells were not detected. Conclusion: Suspended cell spheres generated from OE-19 esophageal carcinoma cells in serum-free stem medium are enriched in tumor stem-like cells. CD44 may be a marker for these cells.

Fundamental Cultural Characteristics of Chlorociboria spp. Native to Korea (한국에 자생하는 녹청균류의 기초 배양 특성)

  • Jeon, Sung-Min;Ka, Kang-Hyeon;Wang, Eun-Jin;Ryoo, Rhim;Jang, Yeongseon
    • The Korean Journal of Mycology
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    • v.46 no.2
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    • pp.145-160
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    • 2018
  • Members of Chlorociboria are soft-rot ascomycetes that produce blue-green pigment. We investigated the growth characteristics of two Korean species of Chlorociboria, eight strains of Chlorociboria aeruginascens and Chlorociboria poutoensis, under various culture conditions (solid media, temperature, pH) and screened them for extracellular enzyme activity. Although the growth rate was slow, all tested strains of Chlorociboria spp. grew well on potato dextrose agar (PDA; 16.3~42.6 mm after 60 days) or Sabouraud dextrose agar (SDA), but not on malt extract agar (MEA). Compared with C. aeruginascens strains, C. poutoensis strains exhibited higher expression of blue-green pigments on both PDA and SDA media. The optimal temperature for mycelial growth was $20{\sim}25^{\circ}C$, and mycelial growth was lower at $30^{\circ}C$ than at $10^{\circ}C$. All strains tended to have increased mycelial growth as the incubation temperature increased in the range of 10 to $20^{\circ}C$. The optimal pH of potato dextrose broth (PDB) for mycelial growth varied according to the strain under static culture conditions. Maximum biomass production was obtained at pH 6.0 for NIFoS 579 ($114.3{\pm}5.1mg/60days$), but it maintained a stable pigment expression under a broad pH spectrum. The activities of both cellulase and laccase were observed in all tested strains of Chlorociboria spp. Enzyme activities of NIFoS 579 were remarkably higher than those of the other strains. From these results, we suggest that C. poutoensis NIFoS 579 is a potential candidate for use as a source of natural blue-green dye.

TIMP-2 Gene Transfer Via Adenovirus Inhibits the Invasion of Lung Cancer Cell (TIMP-2 유전자 재조합 아데노바이러스의 폐암세포 침윤 억제 효과)

  • Oh, Yeon-Mok;Lee, Jae-Ho;Yoo, Chul-Gyu;Chung, Hee-Soon;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.2
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    • pp.189-197
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    • 2000
  • Background : Tissue inhibitor of metalloproteinase is a natural inhibitor that counteracts pro teolytic enzymes essential to the invasion of cancer cell. Whether or not TIMP-2 gene transfer via adenovirus could inhibit the invasion of lung cancer cell iη vitro was evaluated for the future purpose of gene therapy against lung cancer. Methods : Recombinant adenovirus-TIMP-2(Ad-TIMP-2) was generated by homologous recombination after pACCMV-TIMP-2 and pJM17 were cotransfected into 293 cell by standard calcium phosphate coprecipitate method. Calu-6, one of the most invasive lung cancer cells, was transduced with Ad-TIMP-2 or Ad-$\beta$gal. Anchorage-independent growth and invasiveness were assessed by soft agar clonogenicity assay and invasion assay using two-chamber, well divided by matrigel. Results : Ad-TIMP-2 transduced calu-6 cells produced biologically active TIMP-2 more than 50 times more than parental calu-6. TIMP-2 gene transfer did not suppress the in vitro tumorigenicity. However, two chamber well assay revealed that Ad-TIMP-2 transduction reduced the invasiveness of calu-6 efficiently (12% compared with parental cell) even at low 10moi. Conclusion : Even though TIMP-2 gene transfer did not inhibit in vitro tumorigenicity, it did inhibit invasion of lung cancer cell in vitro. The inhibition of invasion by Ad-TIMP-2 may be a useful strategy for the treatment of lung cancer.

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Occurrence of Rhizopus Soft Rot on Peach (Prunus persica var. vulgaris) Caused by Rhizopus nigricans in Korea

  • Kwon, Jin-Hyeuk;Kang, Soo-Woong;Park, Chang-Seuk
    • Mycobiology
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    • v.28 no.4
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    • pp.177-179
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    • 2000
  • A soft rot of fruits caused by Rhizopus nigricans occurred on peach (Prunus persica var. vulgaris) in The Chinju City Agricultural Products Wholesale Marke during in summer season of 2000. The disease infection usually started from wounding after harvest fruits, and then moved to outside. At first, the lesions started with water soaked and rapidly softened and diseased area gradually expanded. In severely infected film house, the rate of infected fruits reached 65.2%. Numerous sporangiospores were produced on the diseased fruits. Most of the sporangiospores were appeared to be readily dispersed in the air. The mycelia grew surface of fruits and produced stolons. Colonies on potato dextrose agar at $25{\sim}30^{\circ}C$ white cottony at first becoming heavily speckled by the presence of sporangia and the browinish black at maturity, spreading rapidly by means of stolons fired at various points to the substrate by rhizoids. Sporangia were $85.3{\sim}243.5{\times}53.4{\sim}219.2\;{\mu}m$ in size and were globose or sub-globose with. somewhat flattened base. The color of sporangia was white at first and then turned black with many spores, and never over-hanging. Sporangiophores were $8.9{\sim}36.6\;{\mu}m$ in width, smooth-walled, non-septate, light brown, simple, long, arising in groups of $3{\sim}5$ from stolons opposite rhizoids. Sporangiospores was $9.7{\sim}24.8{\times}5.9{\sim}15.8\;{\mu}m$, irregular, round, oval, elongate, angular, and browinish-black streaked. Columella was $70.2{\times}149.7{\mu}m$. brownish gray, and umberella-shaped when dehisced. The causal organism was identified as. Rhizopus nigricans Lind on the basis of the morphiogical characteristics of the fungus. Rhizopus soft rot on peach (Prunus persica) caused by the fungi has not been reported in Korea. This is the first report of rhizopus soft rot on peach caused by Rhizopus nigricans in Korea.

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A Basic Study on the Variation of Temperature Characteristics for Attenuation Coefficient and Sound Velocity in Biological Tissues

  • Park, Heung-Ho
    • Journal of Biomedical Engineering Research
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    • v.14 no.3
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    • pp.273-282
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    • 1993
  • This study is concerned with the temperature dependence characteristics of ultrasound parameters in biological tissues, which are basic on the noninvasive deep body temperature estimation. Used parameters are ultrasonic attenuation coefficient and sound velocity In order to accomplishment our purpose, several signal processing methods were used. Attenua4iorl coefficient was estimated by spectral difference method and sound velocity was estimated by P-P method. And we also examined these methods through a series of IN VITRO experi mentis that used tissue-mimicking phantom samples and biological tissue samples. In order to imitate the biological soft tissue two kinds of phantom samples are used, one is agar phantom sample which is composed of agar, graphite, N-propyl alcohol and distilled water, and the other is fat phantom sample which is composed of pure animal fat. And the ultrasound transmission mode and reflection mode experiments are performed on the pig's spleen, kidney and fat. As a result, it is found that the temperature characteristics are uniform in case of phan- tom samples but not in biological tissues because of complicate wave propagation within them. Consequently, the possibility of temperature measurement using ultrasound on biological tissue is confirmed and its results may contribute to the establishment of reference values of internal temperature measurement of biological tissues.

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Selection of High Tobramycin-Producing Mutants (Tobramycin 고생산성 변이주의 분리)

  • 나규흠;김학주;김기태;양중익;김계원
    • Microbiology and Biotechnology Letters
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    • v.19 no.4
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    • pp.343-347
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    • 1991
  • An improved method for the selection of high tobramycin-producing mutants of Streptomyces tenebrarius ATCC 17920 was investigated. By the use of apramycin-containing media, low nebramycin-producing mutants were eliminated. Strain No. 23, resistant to apramycin and kanamycin B and sensitive to tobramycin, was isolated from soils, identified as Pseudomonas paucimobilis and used as a test organism for overlaying the mutants on agar plate. While inhibition zones were not shown when the parent strains were overlaid with soft agar containing the strain No. 23, clear zones were shown when high tobramycin-producing mutants were overlaid. Using this screening strategy, 58 mutants showing clear zones had been isolated. antibiotic activities of which were incresed to 3~8 fold compared to that of parent strain.

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Hydrocolloids Decrease the Digestibility of Corn Starch, Soy Protein, and Skim Milk and the Antioxidant Capacity of Grape Juice

  • Yi, Yue;Jeon, Hyeong-Ju;Yoon, Sun;Lee, Seung-Min
    • Preventive Nutrition and Food Science
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    • v.20 no.4
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    • pp.276-283
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    • 2015
  • Hydrocolloids have many applications in foods including their use in dysphagia diets. We aimed to evaluate whether hydrocolloids in foods affect the digestibility of starch and protein, and their effects on antioxidant capacity. The thickening hydrocolloids: locust bean gum and carboxymethyl cellulose, and the gel-forming agents: agar agar, konjacglucomannan, and Hot & Soft Plus were blended with corn starch and soy protein, skim milk, or grape juice and were examined for their in vitro-digestability by comparing the reducing sugar and trichloroacetic acid (TCA)-soluble peptide, for antioxidant capacity by total polyphenol contents and the 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity. The hydrocolloids resulted in a decrease in starch digestibility with the gel-forming agents. Hydrocolloids diminished TCA-soluble peptides in skim milk compared to soy protein with the exception of locust bean gum and decreased free radical scavenging capacities and total phenolic contents in grape juice. Our findings may provide evidence for the use of hydrocolloids for people at risk of nutritional deficiencies such as dysphagia patients.