• Title/Summary/Keyword: X-cells

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A Role of Cell Adhesion Molecules and Gelatinases in Human Serum-Induced Aggregation of Human Eyelid-Derived Stem Cells In Vitro

  • Yang, Hyejin;Lim, Yoon Hwa;Yun, Sujin;Yoon, A Young;Kim, Haekwon
    • Development and Reproduction
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    • v.17 no.4
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    • pp.409-420
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    • 2013
  • Human serum (HS) has been reported to induce aggregation of human eyelid adipose-derived stem cells (HEACs) during high-density culture in vitro. The present study focused on the role of cell adhesion molecules and gelatinases during HS-induced aggregation of HEACs. HS-induced aggregation occurred between 9-15 days of culture. Cells aggregated by HS medium (HS-agg) showed stronger expression of ${\alpha}2$, ${\alpha}2B$, ${\alpha}X$, and CEACAM1 genes compared to non-aggregated cells in HS medium (HS-ex) or in control FBS-cultured cells. HS-agg were distinctly labeled with antibodies against ${\alpha}2$, ${\alpha}2B$, and ${\alpha}X$ proteins. Western blot results demonstrated that the two integrin proteins were greatly expressed in HS-agg compared to HS-ex and control FBS-cultured cells. Treatment of HEACs with anti-integrin ${\alpha}2$ antibody during culture in HS medium delayed aggregation formation. HS-agg exhibited strong expression of MMP1 and MMP9 compared to HS-ex or FBS-cultured cells. Conditioned media from HS-culture showed remarkable increase of MMP9 gelatinolytic activity in comparison to those from FBS-culture. However, there was no change of TIMP mRNA expression in relation to the HS-induced aggregation. Based on these results, it is suggested that integrin ${\alpha}2$, ${\alpha}2B$, and ${\alpha}X$, and MMP9 might play an important role in the HS-induced aggregation of HEACs.

Extracellular Concentration of ⳑ-Cystine Determines the Sensitivity to System xc- Inhibitors

  • Abdullah, Md;Lee, Seung Jin
    • Biomolecules & Therapeutics
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    • v.30 no.2
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    • pp.184-190
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    • 2022
  • Targeting the cystine/glutamate exchange transporter, system xc-, is a promising anticancer strategy that induces ferroptosis, which is a distinct form of cell death mediated by iron-dependent lipid peroxidation. The concentration of ⳑ-cystine in culture medium is higher than the physiological level. This study was aimed to evaluate the effects of ⳑ-cystine concentration on the efficacy of ferroptosis inducers in hepatocellular carcinoma cells. This study showed that treatment with sulfasalazine or erastin, a system xc- inhibitor, decreased the viability of Huh6 and Huh7 cells in a dose-dependent manner, and the degree of growth inhibition was greater in medium containing a physiological ⳑ-cystine concentration of 83 µM than in commercial medium with a concentration of 200 µM ⳑ-cystine. However, RSL3, a glutathione peroxidase 4 inhibitor, decreased cell viability to a similar extent in media containing both ⳑ-cystine concentrations. Sulfasalazine and erastin significantly increased the percentages of propidium iodide-positive cells in media with 83 µM ⳑ-cystine, but not in media with 200 µM ⳑ-cystine. Sulfasalazine- or erastin-induced accumulation of lipid peroxidation as monitored by C11-BODIPY probe was higher in media with 83 µM ⳑ-cystine than in media with 200 µM ⳑ-cystine. In contrast, the changes in the percentages of propidium iodide-positive cells and lipid peroxidation by RSL3 were similar in both media. These results showed that sulfasalazine and erastin, but not RSL3, were efficacious under conditions of physiological ⳑ-cystine concentration, suggesting that medium conditions would be crucial for the design of a bioassay for system xc- inhibitors.

Direct Evidence for the Radioprotective Effect of Various Carbohydrates on Plasmid DNA and Escherichia coli Cells

  • Ryu, Hwa-Ja;Yi, Kyung-Eun;Kim, Do-Won;Jung, Yun-Dae;Chang, Suk-Sang;Seo, Eun-Seong;Lee, Ki-Young;Marceau-Day, M.L.;Kim, Do-Man
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.598-602
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    • 2002
  • Damage to cells exposed to radiation is primarily attributed to direct effects on the structure of cellular DNA. Radiation-induced damage of pBluescript SK plasmid DNA and Escherichia coli $DH5\alpha$ were examined in the presence of various branched oligosaccharides, polysaccharides, and/or 8-MOP (8-methoxypsoralen). Branched oligosaccharides efficiently protected DNA and cells exposed to ultrasoft X-ray and UV irradiation. In the presence of 0.2% (w/v) branched oligosaccharides and polysaccharides, DNA can be protected from damage due to W and ultrasoft X-ray by a factor of 1.3-2.1 fo1d and 3.2-8.3 fold, respectively. The protective effect of cells exposed to UV or ultrasoft X-ray was also observed by branched oligosaccharides. The combination of MOP, a photoreagent, with carbohydrates increased the protective effects for DNA and cells, compared with that of a single use of MOP or carbohydrate alone.

Economic Production of $\gamma$-Interferon from Recombinant Human Cells in Serum Free Medium by a Moving Aeration Membrane Bioreactor (교반형 막 반응기를 이용한 재조합 인간 세포의 무혈청 배지에 의한 $\gamma$-Interferon의 생산)

  • Park, Young-Shik;Kim, Hyun-Kyu;Lim, Seo-Kyu;Park, Kyung-You;Lee, Hyeon-Yong
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.389-394
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    • 1994
  • 8 X 10$^{6}$(viable cells/ml) of maximum cell density and 9000(IU/ml) of $\gamma$-IFN production were obtained at 55(ml/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high comsumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decrea- sed, maintaing 1.5 X 10$^{-3}$(IU/cell/day) specific IFN production rate. The cumulatvie $\gamma$-IFN production was 7.2 X 10$^{8}$(IU) for 70 days of the cultivation, which yields 1 X 10$^{7}$ (IU/day) of IFN production rate. Therefore, a commercial production of $\gamma$-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.

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Permeability of (SiO2)1-x(ZnO)x Inorganic Composite Thin Films Deposited as a Passivation Layer of Ca Cell (Ca Cell의 보호막으로 증착된 (SiO2)1-x(ZnO)x 무기 혼합 박막들의 투습 특성)

  • Kim, Hwa-Min;Ryu, Sung-Won;Sohn, Sun-Young
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.22 no.3
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    • pp.262-268
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    • 2009
  • We investigated the properties of inorganic diatomic films like silicon oxide ($SiO_2$) and zinc oxide (ZnO) and their composite films are packed as a passivation layer around Ca cells on glass substrates by using an electron-beam evaporation technique and rf-magnetron sputtering method. When these Ca cells are exposed to an ambient atmosphere, the water vapor penetrating through the passivation layers is adsorbed in the Ca cells, resulting in a gradual progress of transparency in the Ca cells, which can be represented by changes of the optical transmittance in the visible range. Compared with the saturation times for the Ca cells to become completely transparent in the atmosphere, the protection effects against permeation of water vapor are estimated for various passivation films. The thin composite films consist of$SiO_2$ and ZnO are found to show a superior protection effect from water vapor permeation compared with diatomic inorganic films like $SiO_2$ and ZnO. Also, this inorganic thin composite films are also found that their protection effect against permeation of water vapor can be significantly enhanced by choosing their suitable composition ratio and deposition method, in addition, the main factors affecting the permeation of water vapor through the oxide films are found to be the polarizability and the packing density.

A Study on the Effect of "ADAPTAGEN"$^{\textregistered}$ Korean Ginseng Components, for the Injured Mouse by X-ray($^{60}Co$) Irradiation (X-방사선($^{60}Co$)에 조사된 새앙쥐의 상해에 대한 "아답태겐"$^{\textregistered}$의 효과에 관한 연구)

  • 공태훈;유성렬
    • Journal of Ginseng Research
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    • v.15 no.3
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    • pp.171-178
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    • 1991
  • The results ok feeding experiments to the mice with ginseng extract, ginseng Powder, and ADAPTAGEN, for 30 days before X-ray irradiation and for 40 days after the X-ray irradiation at 750 rads were as follow: 1. The 50% lethals days (LD50, ) by the X-ray irradiation were 9 days at 1, 000 rads. 10 days at 900 rads, 11 days at 800 rads, 14 days at 760 rads, and 19 darts at 750 rads. Therefore, the standard radiation dose was set at 750 radb/8 min. 2. The 80% of the control group mice exposed to the X-ray radiation without ginseng feeding died in periods ranging from 14 to 24 days and the 20~30% of the ginseng extract and ginseng powder feeding groups died. But the 100% of the mice fed with ADAPTAGEN survived. 3. Testicles of the control group became smaller in weight than the nomad group by 26.5 to 29.0% and those of the ginseng extract and ginseng powder feeding group reduced by 44.6 to 60.4%. However, testicles of the ADAPTiIGEN feeding group increased in size by 77.4% to 87.1% and in weight by 61%, showing a recovery phenomenon approarhing to those of the ordinary mice. The ADAPTAGEN feeding group mice were also as active in color as the ordinary ones. 4. An electron micrograph(X8, 000X2.2) of the liver cells of the mice which had been 40 days after X-ray irradiation showed as follows; The control group appeared that is physiological action stopped due to the frequent occurrence of morphological change of the nucleus and diffusion of chromosome, reduction in microspores and expansion of microsomts, and endoplasmic change of mitochondria. The liver cells or the ADAPTAGEN feeding group were in a state similar to those of the ordinary mice restoring to normalcy In contrast, the liver cells of the ginseng extract and ginseng powder feeding groups were still far from being normal. 5. A serological analysis showed that the control group sharply decreased in albumin, Y-g1obu1in, and IgG so far as to cause dystrophy and to weaken antibody resistance but that ginseng extract and ginseng powder feeding groups, though in a little more restoring state than the control group, were still far from the normal group. The ADAPTAGEN feeding group restored to a state as comparable to the normal group in the contents of albumin ${\gamma}$-globulin, IgG and serum protein. In order words, it is noteworthy that ADAPTAGEN feeding was effective in revitalizing the destroyed cells of a living body and that it has the function of normalizing antibody components.

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Adipose-Derived Stem Cells Suppress Inflammation Induced by IL-1β through Down-Regulation of P2X7R Mediated by miR-373 in Chondrocytes of Osteoarthritis

  • Jin, Rilong;Shen, Miaoda;Yu, Liedao;Wang, Xuanwei;Lin, Xiangjin
    • Molecules and Cells
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    • v.40 no.3
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    • pp.222-229
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    • 2017
  • Adipose-derived stem cells (ADSCs) were previously considered to have an anti-inflammatory effect, and Interleukin-$1{\beta}$ ($IL-1{\beta}$) was found to be a pro-inflammatory factor in chondrocytes, but the mechanism underlying ADSCs and $IL-1{\beta}$ is unclear. In this study, we investigate whether P2X7 receptor (P2X7R) signalling, regulated by microRNA 373 (miR-373), was involved in the ADSCs and $IL-1{\beta}$ mediated inflammation in osteoarthritis (OA). Chondrocytes were collected from 20 OA patients and 20 control participants, and ADSCs were collected from patients who had undergone abdominal surgery. The typical surface molecules of ASDCs were detected by flow cytometry. The level of nitric oxide (NO) was determined by Griess reagent. Concentrations of prostaglandin E2 (PGE2), interleukin 6 (IL-6), matrix metallopeptidase 3 (MMP-3) were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of IL-6, MMP-3, miR-373 and P2X7R were determined by real-time polymerase chain reaction (PCR), and Western blot was used to detect the protein expression of P2X7R. The typical potential characters of ADSCs were verified. In chondrocytes or OA tissues, the miR-373 expression level was decreased, but the P2X7R expression was increased. $IL-1{\beta}$ stimulation increased the level of inflammatory factors in OA chondrocytes, and ADSCs co-cultured with $IL-1{\beta}$-stimulated chondrocytes decreased the inflammation. OA chondrocytes transfected with the miR-373 inhibitor increased the inflammation level. The miR-373 mimic suppressed the inflammation by targeting P2X7R and regulated its expression, while its effect was reversed by overexpression of P2X7R. $IL-1{\beta}$ induced inflammation in OA chondrocytes, while ADSCs seemed to inhibit the expression of P2X7R that was regulated by miR-373 and involved in the anti-inflammatory process in OA.

Morphological Study on the TUNEL Reaction of Mouse Ovary by X-ray Irradiation (방사선 조사선량에 따른 생쥐 난소의 TUNEL 염색반응에 관한 형태학적 연구)

  • Yoon, Chul-Ho;Chang, Byung-Soo
    • Applied Microscopy
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    • v.37 no.2
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    • pp.135-142
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    • 2007
  • This research investigated morphological changes of ovarian follicle according to dose of irradiation when adult mice were exposed to X-rays from 6 MeV LINAC. At day 3 after irradiation of 200 cGy, 400 cGy and 600 cGy X-ray to the hole body of mice, the ovaries collected and stained with TUNEL. The normal follicles and atretic follicles were identified to apoptosis by the staining with TUNEL. In the atretic follicles of the normal ovary, the apoptotic bodies were well appeared and stained brown color. Almost of the follicles following irradiation are stained with TUNEL, but the sensitivity of reaction is weaker than that in irradiation of 400 cGy and 600 cGy X-ray. The granulosa cells of the radiated normal follicle by 400 cGV are shown brown color. In this stage, the nucleus of granulosa cells in the atrectic follicles are condensed and picknotic feature. The size of the radiated follicle by 600 cGy are decreased than the normal follicles. The atropic follicles are filled with apoptotic bodies which change of granulosa cells and theca cells by influence of X-ray. All of cell in the follicles are strongly positive stained with TUNEL by irradiation of 600 cGy.

Studies on the Estimation of Pregnancy Rate of White Hanwoo (Albino Korean Native Cattle) with Sexed Semen (백한우의 성판별 정액을 이용한 수태율 추정에 관한 연구)

  • Kim, Sung Woo;Choi, Jinseok;Choe, Changyoung;Kim, Dongkyo;Ko, Yeoung-Gyu;So, Chungsil;Seong, Hwan-Hoo
    • Journal of Embryo Transfer
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    • v.30 no.1
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    • pp.45-50
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    • 2015
  • The purpose of this study is to produce wanted sex progeny of genetically confined White Hanwoo (albinism) with preselected sex sperm. One bull of White Hanwoo was chosen for semen donor and X sperm was sorted by MoFlo XDP cell sorter. To compare the pregnancy and birth rates, KPN straw was used as control, total number of unsorted sperm was $20{\times}10^6/straw$. Sexed X frozen semen with $20{\times}10^6$ cells or $4{\times}10^6$ cells per straw were in seminated twice on Hanwoo heifers. The abnormality of the sexed X semen was $24.9{\pm}7.31%$ and distal reflex abnormality of mid piece was significantly (p<0.05) higher (11.7%) compared with that of KPN 768 (5.6%). There were no differences on the pregnancy and birth rates between $2{\times}10^6$ cells or $4{\times}10^6$ cells of X-sperm but KPN semen showed significant differences (p<0.05). The pregnancy rates of KPN 768, $2{\times}10^6$ cells and $4{\times}10^6$ cells X-sperm of White Hanwoo cattle were 85.0%, 26.3% and 50%. The birth rates were 80.0%, 15.8% and 21.4%, respectively. The female offspring rates of KPN 768, $2{\times}10^6$ cells and $4{\times}10^6$ cells X-sperm of White Hanwoo cattle were 43.8%, 100% and 100% (p<0.05). These results indicated that sex sorted White Hanwoo could be used for the production of wanted progeny with $2{\times}10^6$ cells/straw for AI. To increase the efficiency of calf production, the sperm number of sex sorted semen will be optimized for sex selection of White Hanwoo progeny.

The Cell Protective Effects of Dioscoreae Rhizoma by Antioxidant Activities on HeLa Cells (HeLa cell에서 산약(山藥)의 항산화 작용을 통한 세포보호효과에 대한 연구)

  • Yang, Jeong-Min;Jun, Yung-Joon;Nam, Ju-Young;Son, Mi-Young;Sung, Jung-Suk;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.21 no.2
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    • pp.97-107
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    • 2008
  • Purpose: This study is to examine antioxidant activities of Dioscoreae Rhizoma on HeLa cell Methods: Aqueous extract was used to treat HeLa cell with different concentrations treated with water or MeOH extract of Dioscoreae batatas (0, x10, x20, x40, x80). The MTT reduction assay and flow cytometric analysis was employed to quantify the differences in cell activity and viability. Results: Co-treatment with $H_2O_2$ and Dioscoreae batatas extracts reduced apoptosis of HeLa cells by decreasing G2/M arrest. Dioscoreae batatas extracts increased the survival rate of cells treated with cisplatin and Scutellaria barbata. Conclusion: Our results suggest that Dioscoreae Rhizoma extracts induce cell protective effect by antioxidant activities.

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