• Title/Summary/Keyword: 세포생존

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Chromosome and Spindle Configuration of Mouse Oocytes after Vitrification at the Mature Stage (마우스 성숙난자의 유리화 동결법에 따른 동결 융해 후의 염색체와 방추사의 분석)

  • ;;;;Gary B. Anderson
    • Korean Journal of Animal Reproduction
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    • v.25 no.3
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    • pp.287-292
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    • 2001
  • Selection of oocyte cryopreseivation method is a prerequisite factor for developing an effective bank system. To develop an effective vitrification method, we examined whether damages in spindle and chromosome morphology induced by vitrification. Intact cumulus-enclosed oocytes were vitrified with DPBS with 5.5 M ethylene glycol and 1.0 M sucrose, and loaded onto eletron microscopic copper grid for storing in liquid nitrogen. Intact vitrified and thawed oocytes were immunostaining for tubulin and karyotying for chromosome. Vitrfied and thawed oocytes had a higher rate of chromosome (32.8% vs. 19.6%) and spindle (32.3% vs. 20.2%) abnormalities compared with fresh oocytes. Mouse oocytes after vitrification at the mature stage showed increased incidence of chromosomal and spindle abnormalities.

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Viability of cells in aspirated fat tissue after 1 year cryopreservation (1년 이상 냉동 보관한 흡인 지방조직 내의 세포 생존)

  • Son, Daegu;Oh, Jaehoon;Choi, Taehyun;Kim, Junhyung;Han, Kihwan
    • Archives of Plastic Surgery
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    • v.36 no.2
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    • pp.135-139
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    • 2009
  • Purpose: The use of an autogenous fat graft has become a common procedure in plastic surgery. However, questions remain concerning on the viability of fat cells and preservation method of aspirated fat. The purpose of this study was to examine the viability of fat cells stored at $-20^{\circ}C$ in the freeze for 1 year after harvest from abdominal liposuction. Methods: Eighteen adults (aged 24 to 65 years old, 16 female and 2 male) were recruited for this study. Harvested aspirated fat tissues were obtained by suction - assisted lipectomy and frozen at $-20^{\circ}C$ commercial refrigerator for one year (average 12.5 months). The viability off at cells in specimens were measured after thawing. The numbers of viable cells were measured on a fluorescence microscope after staining with fluorescein diacetate and propidium iodide. GPDH (Glycerol - 3 - phosphate dehydrogenase) activity was measured. Cell culture was done for 3 weeks. Results: There were no viable cells under the fluorescence microscope, no detectable GPDH activity, and no cultured cells. Conclusion: These findings suggest that aspirated fat after frozen storage for one year at $-20^{\circ}C$ freezer is inadequate to reuse.

Mass Propagation of Dicentra spectabilis L. Lemaire Through In vitro Suspension Culture (현탁배양을 통한 금낭화(Dicentra spectabilis L. Lemaire)의 대량증식)

  • Lee, Kang-Seop;Sim, Ock-Kyeong;Shin, Jeong-Sun;Choi, Yong-Eui;Kim, Ee-Yup
    • Journal of Plant Biotechnology
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    • v.31 no.2
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    • pp.121-126
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    • 2004
  • Bleeding heart (Dicentra spectabilis L. Lemaire) is one of the most valuable wild flower in Korea. This work was conducted for the mass production of somatic embryos through suspension culture and more effective plant regeneration system in Dicentra spectabilis. High-frequency embryogenic callus proliferation was achieved in SH liquid medium supplemented with 1 mg/L 2,4-D. Half-strength SH medium was suitable concentration for somatic embryo induction and germination. About 5,000 embryos were produced per 250$m\ell$ flask after 4 weeks of culture. Germination rate of somatic embryos was decreased when GA$_3$ was added in medium. The plantlets showed a 58% survival rate when transferred to pots after 1 month of culture. The results indicate that micropropagation procedure via somatic embryogenesis can be applied for an efficient mass propagation of Dicentra spectabilis.

Immunostimulatory effects of BCG-CWS on the proliferation and viability of mouse spleen cells (마우스 비장세포의 증식과 생존율에 대한 BCG-CWS의 면역자극 효과)

  • Lee, Che-Wook;Ko, Eun-Ju;Joo, Hong-Gu
    • Korean Journal of Veterinary Research
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    • v.52 no.2
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    • pp.89-97
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    • 2012
  • Mycobacterial cell-wall skeleton (CWS) is an immunoactive and biodegradable particulate adjuvant and has been tried to use for immunotherapy. The CWS of Mycobacterium bovis bacillus Calmette-Guerin (BCG-CWS) was studied as an universal vaccine vehicle for antigen conjugation, to develop potentially effective and safe vaccine. Although a variety of biological activities of BCG-CWS have been studied, the effects of BCG-CWS on spleen cells are not fully elucidated. Using MTT assay and trypan blue exclusion test, we found that BCG-CWS significantly enhanced the viability and proliferation of cells. Multiple clusters, indicating proliferation, were observed in BCG-CWS-treated spleen cells and surface marker staining assay revealed that BCG-CWS promoted the proliferation of $CD19^+$ B lymphocyte rather than $CD4^+$ or $CD8^+$ T lymphocyte. In addition, BCG-CWS up-regulated the expression of anti-apoptotic molecules such as bcl-2, bcl-xL. BCG-CWS increased the surface expression of CD25 and CD69 as well as IL-2 production of spleen cells, suggesting increased activation. Furthermore, BCG-CWS enhanced the antigen-specific cell proliferation and interferon-gamma production of spleen cells. Taken together, these results demonstrate the immunostimulatory effects of BCG-CWS on spleen cells via multiple mechanisms, providing valuable information to broaden the use of BCG-CWS in clinical and research settings.

Effect of Tea Catechin, EGCG (Epigallocatechin Gallate) on Killing of Oral Bacteria (차 카테킨 EGCG (Epigallocatechin Gallate)의 구강세균에 대한 살균효과)

  • Yu Mi-Ok;Chun Jae-Woo;Oh Kye-Heon
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.364-366
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    • 2004
  • The purpose of this work was to investigate the effect oftea catechin, epigallocatechin gallate (EGCG) on killing of oral bacteria. The antibacterial activity of 2.5 mg/ml and 5.0 mg/ml EGCG was investigated for target bacteria of which initial cell number was approximately adjusted to $10^{7}ml$. The antibacterial activity of EGCG was proportional to the concentration according to colony-forming unit(CFU) of target bacteria enumerating on selective and complex media. Streptococcus mutans and Streptococcus sobrinus at 5mg/ml EGCG were completely killed within 8 hrs. Lactobacillus plantarum and Lactobacillus acidophilus were also killed within 2 hrs and 4 hrs under the same conditions, respectively. Oral bacteria at 2.5 mg/ml EGCG were completely killed within 10 hr. Colony numvers of S. mitis and S. salivarius treated with 2.5 mg/ml EGCG were decreased on MS solid media and no colony was observed on the media within 12 hrs. In consequence, EGCG would be a natural and effective compound that kill oral bacteria being caused of bad breath, plaque and gingivitis, and for preventing and treating dental caries.

Screening and Partial Purification of Haloperoxidase from Marine Actinomycetes (해양방선균으로부터 Haloperoxidase의 검색과 특성)

  • Cho, Ki-Woong
    • Korean Journal of Microbiology
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    • v.44 no.2
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    • pp.116-121
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    • 2008
  • In my search of microbial source of novel enzymes, a marine actinomycetes, A1460, producing haloperoxidase was isolated from macroalgae from south sea, Korea and studied for physiological and biochemical properties. The haloperoxidation reaction was followed by the bromination of phenol red in the presence of hydrogen peroxide and potassium bromide. The haloperoxidase was partially purified from the cell extract with $35\sim75%$ ammonium sulfate precipitation, High-Q anion exchange chromatography, gel filtration chromatography, hydroxyapetite chromatography and hydrophobic interaction chromatography to a yield of 42% and purification fold of 70. This enzyme showed relatively high heat stability without losing 50% of activity after 1 hr incubation at $60^{\circ}C$. The highest activity was found at $45^{\circ}C$, and the optimal pH was about pH 7, but higher stability was observed at pH 8. Azide and cyanide ion showed strong inhibition at less than 1 $\mu M$ level suggesting that the enzyme was Fe ion dependent haloperoxidase.

Synergistic Effects of Ionizing Radiation and Mercury Chloride on Cell Viability in Fish Hepatoma Cells (이온화 방사선 및 염화수은 처리에 따른 어류 간암세포의 생존능 평가)

  • Han, Min;Hyun, Kyung-Man;Nili, Mohammad;Hwang, In-Young;Kim, Jin-Kyu
    • Korean Journal of Environmental Biology
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    • v.27 no.2
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    • pp.140-145
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    • 2009
  • All organisms are being exposed to harmful factors present in the environmental. The combined action of various factors is a distinguishing feature of modern life. An interaction between two chemicals is considered as synergistic when the effect produced is greater than the sum of the two single responses. The biological effects due to the combined action of ionizing radiation with the other factor are hard to estimate and predict in advance. In the current study, we investigated the synergistic effects between ionizing and $HgCl_2$ using fish hepatoma cells (PLHC-1 cells). The results showed a dramatic decrease of cell viability after simultaneous treatment of PLHC-1 cells with ionizing radiation and $HgCl_2$. Neiither of the two had any cytotoxic effect when treated alone. The cytotoxicity of ionizing radiation was enhanced in the presence of $HgCl_2$. The synergistic effects were observed after exposure of the PLHC-1 cells to ionizing radiation combined with $HgCl_2$. The synergistic interaction was due to an increase of irreversibly damaged cells after the combined exposure. Analysis of the extent of synergistic interaction enables to make quantitative estimation of irreversibly damaged cells after the combined exposure. The present study suggests that PLHC-1 cells can serve as rapid screening tools for detecting the toxicity of harmful factors.

Antioxidantive Effectiveness of Trichosanthes kirilowii Maximowicz Extracts (하늘타리(Trichosantes kirilowii Maximowicz)추출물의 항산화 효과)

  • Zhoh, Choon Koo;Uhm, Tae Yong;Kim, Joo Chan
    • Applied Chemistry for Engineering
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    • v.18 no.6
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    • pp.625-629
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    • 2007
  • Recently, there is a growing interest about unsaturated lecithin having excellent characteristics of skin affinity and absorbency. Accordingly, this study intended to develop a natural sulfuration material in order to enhance the stability of oxidation of unsaturated lecithin and substitute existing sulfuration materials which indicate variability and toxicity. As sulfuration components, plenol acid family, 3,5-dihydroxybenzoic acid, and flavanone were analyzed. Total polyphenol content was higher in the root extracts (133.85 mg/g) than in the fruit extracts (53.5 mg/g). Above 100 ppm polyphenol content, the free radical removal efficiency and lipid oxidation prevention of the root extracts were 20.1 and 19.2% superior compared with BHT respectively. Also, the extracts indicated high survival rate of more than 95% below 1250 ppm, showing the stability. For the stability of liposome made from an unsaturated lecithin, the root extracts were superior to the fruit extracts. Especially, 15.1 and 13.9% of sulfuration effect and zeta potential were improved with 9.3% reduced particle size compared with BHT as the control group, respectively.

Utilization of Supplemental Kluyveromyces fragilis, Candida utilis, Saccharomyces cerevisiae or Brewer제s Yeast in the Formulated Diets for Juvenile Abalone (Halioties discus hannai) (참전복 배합사료의 첨가제로서 Kluyeromyces fragilis, Candida utilis, 빵효모 및 맥주효모의 이용성)

  • 이상민;김동주;김중균;이종관;박상언
    • Journal of Aquaculture
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    • v.13 no.1
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    • pp.55-62
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    • 2000
  • A 10-week feeding trial was conducted to investigate the effects of several yeasts with or without chemical treatment (protoplasted) in formulated diets on growth and body composition of juvenile abalone(Haliotis discus hanai). There replicate groups of the abalone average weighing 210 mg were fed one of eight isonitrogenous (30%) and isolipidic (4.4%) diets containing 3% Kluyveromyces fragilis protoplasted K. fragilis Candida utilis protoplasted C. utilis, Saccharomyces cerevisiae protoplasted S. cerevisiae or brewer's yeast. In addition these formulated diets were compared with commercial diet. Survival rate and proximate analysis of soft whole body of abalone were not significantly affected by the different dietary yeasts and commercial diet (P>0.05) Body weight gain and soft body weight control diet and diets containing S. cerevisiae or brewer's yeast (P<0.05) Shell length of abalone fed yeast and commercial diet (P<0.05) Th results suggest that protoplasted K. fragilis as an additive in this formulated diet can improve weight gain of abalone.

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Anti-aging Effect of Amino Acid Complex on the Skin (Amino Acid Complex의 항노화 작용)

  • Kim, Ki-Ho;Kim, Ki-Soo;Kim, Young-Heui;Ko, Kang-Il;Park, Sun-Hee;Kim, Jin-Guk;Chio, Hyun-Jin;Ko, Su-Yeon;Bae, Yoon-Joo;Kim, Yong-Min
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.2 s.57
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    • pp.81-88
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    • 2006
  • Amino acid complex was made with the composition of amino acids quite similar to that of stratum corneum on the skin. In order to evaluate the efficacy of amino acid complex on the skin as an active cosmetic ingredient, we measured cytotoxicity. TIMP-1 expression, moisturizing effect and the amount of horny substance. In cytotoxicity assay, amino acid complex did not show any cytotoxicity. In moisturizing effect test using corneometer, it showed very good moisturizing effect. In TIMP-1 mRNA assay using RT-PCR, moo acid complex showed the increase of TIMP-1 expression, suggesting that amino acid complex have anti-wrinkle effect. Therefore, amino acid complex may be useful as an active ingredient for anti-aging.