• Title/Summary/Keyword: Conditioned Medium

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APPLICATION OF HF COASTAL OCEAN RADAR TO TSUNAMI OBSERVATIONS

  • Heron, Mal;Prytz, Arnstein;Heron, Scott;Helzel, Thomas;Schlick, Thomas;Greenslade, Diana;Schulz, Eric
    • Proceedings of the KSRS Conference
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    • v.1
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    • pp.34-37
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    • 2006
  • When tsunami waves propagate across open ocean they are steered by Coriolis force and refraction due to gentle gradients in the bathymetry on scales longer than the wavelength. When the wave encounters steep gradients at the edges of continental shelves and at the coast, the wave becomes non-linear and conservation of momentum produces squirts of surface current at the head of submerged canyons and in coastal bays. HF coastal ocean radar is well-conditioned to observe the current bursts at the edge of the continental shelf and give a warning of 40 minutes to 2 hours when the shelf is 50-200km wide. The period of tsunami waves is invariant over changes in bathymetry and is in the range 2-30 minutes. Wavelengths for tsunamis (in 500-3000 m depth) are in the range 8.5 to over 200 km and on a shelf where the depth is about 50 m (as in the Great Barrier Reef) the wavelengths are in the range 2.5 - 30 km. It is shown that the phased array HF ocean surface radar being deployed in the Great Barrier Reef (GBR) and operating in a routine way for mapping surface currents, can resolve surface current squirts from tsunamis in the wave period range 20-30 minutes and in the wavelength range greater than about 6 km. There is a trade-off between resolution of surface current speed and time resolution. If the radar is actively managed with automatic intervention during a tsunami alert period (triggered from the global seismic network) then it is estimated that the time resolution of the GBR radar may be reduced to about 2 minutes, which corresponds to a capability to detect tsunamis at the shelf edge in the period range 5-30 minutes. It is estimated that the lower limit of squirt velocity detection at the shelf edge would correspond to a tsunami with water elevation of less than 5 cm in the open ocean. This means that the GBR HF radar is well-conditioned for use as a monitor of small and medium scale tsunamis, and has the potential to contribute to the understanding of tsunami genesis research.

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Fertilizing Ability of Bovine Spermatozoa Following Oviduct Epithelial Cell Co-culture In Vitro (난관상피세포와 공배양한 소 정자의 체외수정능)

  • 황우석;노상호;이병천
    • Journal of Embryo Transfer
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    • v.13 no.3
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    • pp.227-233
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    • 1998
  • The aim of these experiments was to investigate the effects of oviduct epithelial cells on bovine in vitro fertilization. Oviduct epithelial cell monolayers (OEC) on the 4-well dish were prepared according to general procedures. Monolayers were formed within 5days. The medium for OEC culture (TCM199 with 10% FBS) was replaced with IVF-TALP 2h before each experiment. Macromolecules/proteins from oviductal conditioned medium (OM) were recovered by ultrafiltration, which desalted and concentrated macromolecules greater than 5kDa, and this OM were added to W medium (experiment 1). The cleavage rate in OM+OEC group was significantly higher than in OM group (p〈0.01). In this experiment 2, oocytes were inseminated on OEC with sperm which had been pre-incubated with OEC for 0 or 4h before insemination. In this experiment, oocytes were exposed to sperm only 8 h for clarifying the effect. After insemination, oocytes were cultured in CRlaa. At 42 h post insemination, oocytes were denuded and examined for evidence of cleavage. The cleavage rates of oocytes which were inseminated with OEC treated sperm for 4 h were significantly higher than those of the other group (p〈0.01). In conclusion, sperm released from OEC have more fertilizing ability than those before attachment.

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The Study on Mucin Release by Airway Goblet Cells in Primary Culture

  • Yang, Ji-Sun;Kim, Ok-Hee;Roh, Yong-Nam;Yi, Sook-Young;Park, Ki-Hwan;Rheu, Hang-Mook
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.89-89
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    • 1995
  • Surface epithelial cells isolated from hamster tracheas and grown on a thick collagen gel become a highly enriched population of mucus-secreting cells. Epithelial cells from tracheas of hamsters were collected using enzymatic procedures and cultured under various conditions. The medium used consisted of a 1:1 mixture of medium 199 and Dulbecco's modified Eagle's (DME) medium which was conditioned before use. Insulin, transferrin, hydrocortisone, epidermal growth factor, and extract from bovine hypothalamus were used as supplement. Due to relatively low basal rates of min secretion from in vitro cultures, cultures are generally radiolabeled using $^3$H-glucosamine as a metabolic precursor. The radiolabeled mucinsreleased are quantitated by precipitation with TCA/PTA. Using this cell culture system, we investigated mucin release of goblet cells by altering the media bathing the apical surface of hamster tracheal surface epithelial(HTSE) cells. Acidic media added sulfuric acid caused sigcificant increases in mucin relesse (155${\pm}$20% at pH 4 and 146${\pm}$16% at, pH 5). Ammonium hydroxide also increased mucin release at pH 9.0(156${\pm}$17%) and pH 10(295${\pm}$9%) respectively. This additional mucin release seems to be associated with cell membrane damage as indicated by release of cellular LDH. SP stimulates secretion of mucin in cultured HTSE cells(154${\pm}$16% at 1${\times}$10$\^$-6/M and 165${\pm}$25% at 1${\times}$10$\^$-5/M. PAF at 5${\times}$10$\^$-6/M and 5${\times}$10$\^$-5/M enhanced by HTSE cells in vitro 168${\pm}$34% and 259${\pm}$30% of mucin secretion, respectively. The increase in mucin release by PAF and SP was not secondary to cell damage or necrosis. SP and PAF may be in mediating mucous secretion induced by inflammation irritantion and infection.

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Induction of a Neuronal Phenotype from Human Bone Marrow-Derived Mesenchymal Stem Cells

  • Oh, Soon-Yi;Park, Hwan-Woo;Cho, Jung-Sun;Jung, Hee-Kyung;Lee, Seung-Pyo;Paik, Ki-Suk;Chang, Mi-Sook
    • International Journal of Oral Biology
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    • v.34 no.4
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    • pp.177-183
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    • 2009
  • Human mesenchymal stem cell (hMSCs) isolated from human adult bone marrow have self-renewal capacity and can differentiate into multiple cell types in vitro and in vivo. A number of studies have now demonstrated that MSCs can differentiate into various neuronal populations. Due to their autologous characteristics, replacement therapy using MSCs is considered to be safe and does not involve immunological complications. The basic helix-loop-helix (bHLH) transcription factor Olig2 is necessary for the specification of both oligodendrocytes and motor neurons during vertebrate embryogenesis. To develop an efficient method for inducing neuronal differentiation from MSCs, we attempted to optimize the culture conditions and combination with Olig2 gene overexpression. We observed neuron-like morphological changes in the hMSCs under these induction conditions and examined neuronal marker expression in these cells by RTPCR and immunocytochemistry. Our data demonstrate that the combination of Olig2 overexpression and neuron-specific conditioned medium facilitates the neuronal differentiation of hMSCs in vitro. These results will advance the development of an efficient stem cell-mediated cell therapy for human neurodegenerative diseases.

Improving the Survival and Maintenance of the Undifferentiated State of Cryopreserved Human Embryonic Stem Cells by Extended Incubation with Feeder Cells Overnight before Vitrification (동결에 앞서 시행된 지지세포와의 추가 공배양이 인간 배아줄기세포의 유리화 동결보존 후 생존율과 미분화 유지에 미치는 영향)

  • Cha, Soo-Kyung;Choi, Kyoung-Hee;Shin, Ju-Mi;Park, Kyu-Hyung;Yoon, Tae-Ki;Chung, Hyung-Min;Lee, Dong-Ryul
    • Development and Reproduction
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    • v.12 no.2
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    • pp.141-149
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    • 2008
  • This study was conducted to develop an efficient cryopreservation method of human embryonic stem (ES) cells using vitrification. In an initial experiment, sub-clumps of human ES cells (CHA-hES3 and CHA-hES4) were vitrified using grids after incubation with STO feeder cells for 1 or 16 h (Groups 1-1 and 1-2, respectively). After storage for $2{\sim}4$ months, thawed clumps were re-plated on a fresh feeder layer. The survival rates of warmed CHA-hES3 and CHA-hES4 cells of Group 1-2 were significantly higher than those of the corresponding Group 1-1 cells. In the second experiment, human ES cells were vitrified after incubation with feeder or feeder-conditioned medium (Groups 2-1 to -7). Relative mRNA expression of BM proteins and survival rates were increased following incubation of ES cells with fresh feeder cells for 16 h. In conclusion, increasing of tight adhesion between ES cells by extended incubation with feeder could reduce cryoinjury after vitrifying/warming.

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Induction of Bone Morphogenetic Protein-2 from Gingival Epithelial Cells by Oral Bacteria

  • Kim, Young-Sook;Ji, Suk;Jung, Hong-Moon;Woo, Kyung-Mi;Choi, Young-Nim
    • International Journal of Oral Biology
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    • v.32 no.3
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    • pp.103-107
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    • 2007
  • We hypothesized that plaque-associated bacteria may have a role in maintenance of alveolar bone. To test it, immortalized gingival epithelial HOK-16B cells were co-cultured with live or lysed eight plaque bacterial species and the expression levels of bone morphogenetic protein (BMP)-2 and -4 were examined by real time reverse transcription-polymerase chain reaction. Un-stimulated HOK-16B cells expressed both BMP-2 and -4. Co-culture with plaque bacterial lysates had significant effects on the level of BMP-2 but not on that of BMP-4. Five species including Streptococcus sanguinis, S. gordonii, Veillonella atypica, Porphyromonas gingivalis, and Treponema denticola substantially up-regulated the level of BMP-2. In contrary to the upregulatory effect of lysate, live T. denticola suppressed the expression of BMP-2. In addition, in vitro osteoblastic differentiation assay using C2C12 cells and the conditioned medium of HOK-16B cells confirmed the production of BMPs by gingival epithelial cells and the modulation of BMP expression by the lysates of S. sanguinis and T. denticola. In conclusion, we have shown that plaque bacteria can regulate the expression of BMP-2 by gingival epithelial cells, the physiologic meaning of which needs further investigation.

Cyanidin and Cyanidin-3-O-β-D-glucoside Suppress the Inflammatory Responses of Obese Adipose Tissue by Inhibiting the Release of Chemokines MCP-1 and MRP-2

  • Choe, Mi-Ran;Kang, Ji-Hye;Yoo, Hoon;Yang, Chae-Ha;Kim, Mi-Ok;Yu, Ri-Na;Choe, Suck-Young
    • Preventive Nutrition and Food Science
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    • v.12 no.3
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    • pp.148-153
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    • 2007
  • Obesity-induced inflammation plays a crucial role in obesity-related pathologies such as type II diabetes and atherosclerosis. Adipose tissue macrophages and the cell-derived proinflammatory chemokines are key components in augmenting inflammatory responses in obesity. Anthocyanins such as cyanidin and $cyanidin-3-O-{\beta}-D-glucoside$ (C3G) are known to elicit anti-inflammatory activities by suppressing the production of proinflammatory mediators such as tumor necrosis factor alpha and nitric oxide in LPS-stimulated macrophages. In the present study, we investigated whether cyanidin and C3G have the potential to suppress the inflammatory responses of adipose cells. Cyanidin and C3G not only suppressed the migration of RAW 264.7 macrophages induced by mesenteric adipose tissue-conditioned medium, but also inhibited the activation of the cells to produce inflammatory chemokines such as monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-related protein-2 (MRP-2) in a dose-dependent manner. Cyanidin and C3G also inhibited the release of MCP-1 and MRP-2 from adipocytes and/or macrophages. These findings suggest that cyanidin and C3G may suppress the inflammatory responses of adipose tissue in obesity.

Glutathione is the Major Defensive Mechanism against Oxidative Stress in Human Embryonic Stem Cell

  • 이건섭;이영재;김은영;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.78-78
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    • 2003
  • Embryonic stem (ES) cells, derived from preimplantation embryo, are able to differentiate into various types of cells consisting the whole body, or pluripotency. In contrast, terminally differentiated cells do not usually alter their nature but frequently die or transform if they are exposed to inappropriate external stimulations. In addition to the plasticity, ES cells are expected to be different from terminally differentiated cells in very many ways, such as patterns of gene expressions, ability and response of the cells in confronting environmental stimulations, metabolism, and growth rate. As a model system to differentiate these two types of cells, human ES cells (MB03) and terminally differentiated cells (HeLa), we examined the ability of these two types of cells in confronting a severe oxidative insult, that is $H_2O$$_2$. Approximately 1$\times$10$^4$ cells were plated in 96 well plate and serum starved for overnight. The conditioned cells were exposed to a various concentration of $H_2O$$_2$ fur 24 hrs and loaded with neutral red (50$\mu\textrm{g}$/ml) for 4 hrs, washed with PBS for 2 min three times, and entrapped dye was dissolved out using acetic ethanol. Cytotoxicity was determined by reading the amount of dye in the medium using microplate reader. equipped with 575 nm filter. Relative amount of the dye entrapped within MB03 or HeLa were not significantly different when cells were exposed up to 0.4 mM $H_2O$$_2$. However, this sharply decreased down to 0.12% in HeLa cells when the cells were exposed to 0.8 mM $H_2O$$_2$, while it was approximately 54% in MB03 suggesting that this concentration of $H_2O$$_2$ is the defensive threshold for HeLa cells. The resistance to oxidative stimulation reversed, however, when cells were co-treated with BSO (L-buthionine- 〔S, R〕-sulfoximine) which chelates intracellular GSH. This result suggests that cellular GSH is the major defensive mechanism of human ES cells. Induction of enzymes involved in GSH metabolism and type of cell death is currently being studied.

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Quantitative Changes in Tumor-Associated M2 Macrophages Characterize Cholangiocarcinoma and their Association with Metastasis

  • Thanee, Malinee;Loilome, Watcharin;Techasen, Anchalee;Namwat, Nisana;Boonmars, Thidarut;Pairojkul, Chawalit;Yongvanit, Puangrat
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.7
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    • pp.3043-3050
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    • 2015
  • The tumor microenvironment (TME) includes numerous non-neoplastic cells such as leukocytes and fibroblasts that surround the neoplasm and influence its growth. Tumor-associated macrophages (TAMs) and cancerassociated fibroblasts (CAFs) are documented as key players in facilitating cancer appearance and progression. Alteration of the macrophage (CD68, CD163) and fibroblast (${\alpha}-SMA$, FSP-1) cells in Opisthorchis viverrini (Ov) -induced cholangiocarcinoma (CCA) was here assessed using liver tissues from an established hamster model and from 43 human cases using immunohistochemistry. We further investigated whether M2-activated TAMs influence CCA cell migration ability by wound healing assay and Western blot analysis. Macrophages and fibroblasts change their phenotypes to M2-TAMs (CD68+, CD163+) and CAFs (${\alpha}-SMA+$, FSP-1+), respectively in the early stages of carcinogenesis. Interestingly, a high density of the M2-TAMs CCA in patients is significantly associated with the presence of extrahepatic metastases (p=0.021). Similarly, CD163+ CCA cells are correlated with metastases (p=0.002), and they may be representative of an epithelial-to-mesenchymal transition (EMT) with increased metastatic activity. We further showed that M2-TAM conditioned medium can induce CCA cell migration as well as increase N-cadherin expression (mesenchymal marker). The present work revealed that significant TME changes occur at an early stage of Ov-induced carcinogenesis and that M2-TAMs are key factors contributing to CCA metastasis, possibly via EMT processes.

In vivo anti-metastatic action of Ginseng Saponins is based on their intestinal bacterial metabolites after oral administration

  • Saiki, Ikuo
    • Journal of Ginseng Research
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    • v.31 no.1
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    • pp.1-13
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    • 2007
  • We found that the main bacterial metabolite M1 is an active component of orally administered protopanxadiol-type ginsenosides, and that the anti-metastatic effect by oral administration of ginsenosides may be primarily mediated through the inhibition of tumor invasion, migration and growth of tumor cells by their metabolite M1. Pharmacokinetic study after oral administration of ginsenoside Rb1 revealed that M1 was detected in serum for 24 h by HPLC analysis but Rb1 was not detected. M1, with anti-metastatic property, inhibited the proliferation of murine and human tumor cells in a time- and concentration-dependent manner in vitro, and also induced apoptotic cell death (the ladder fragmentation of the extracted DNA). The induction of apoptosis by M1 involved the up-regulation of the cyclin-dependent kinase(CDK) inhibitor $p27^{Kip1}$ as well as the down-regulation of a proto-oncogene product c-Myc and cyclin D1 in a time-dependent manner. Thus, M1 might cause the cell-cycle arrest (G1 phase arrest) in honor cells through the up/down-regulation of these cell-growth related molecules, and consequently induce apoptosis. The nucleosomal distribution of fluorescence-labeled M1 suggests that the modification of these molecules is induced by transcriptional regulation. Tumor-induced angiogenesis (neovascularization) is one of the most important events concerning tumor growth and metastasis. Neovascularization toward and into tumor is a crucial step for the delivery of nutrition and oxygen to tumors, and also functions as the metastatic pathway to distant organs. M1 inhibited the tube-like formation of hepatic sinusoidal endothelial (HSE) cells induced by the conditioned medium of colon 26-L5 cells in a concentration-dependent manner. However, M1 at the concentrations used in this study did not affect the growth of HSE cells in vitro.