• Title/Summary/Keyword: MDCK cell

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Comparison of Caco-2 and MDCK Cells As an In-Vitro ADME Screening Model (In-Vitro 흡수특성 검색모델로서 Caco-2 및 MDCK 세포배양계의 특성 비교 평가)

  • Go, Woon-Jung;Cheon, Eun-Pa;Han, Hyo-Kyung
    • Journal of Pharmaceutical Investigation
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    • v.38 no.3
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    • pp.183-189
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    • 2008
  • The present study compared the feasibility of Caco-2 and MDCK cells as an efficient in-vitro model for the drug classification based on Biopharmaceutics Classification System (BCS) as well as an in-vitro model for drug interactions mediated by P-gp inhibition or P-gp induction. Thirteen model drugs were selected to cover BCS Class I{\sim}IV$ and their membrane permeability values were evaluated in both Caco-2 and MDCK cells. P-gp inhibition studies were conducted by using vinblastine and verapamil in MDCK cells. P-gp induction studies were also performed in MDCK cells using rifampin and the P-gp expression level was determined by western blot analysis. Compared to Caco-2 cells, MDCK cells required shorter period of time to culture cells before running the transport study. Both Caco-2 and MDCK cells exhibited the same rank order relationship between in-vitro permeability values and human permeability values of all tested model compounds, implying that those in-vitro models may be useful in the prediction of human permeability (rank order) of new chemical entities at the early drug discovery stage. However, in the case of BCS drug classification, Caco-2 cells appeared to be more suitable than MDCK cells. P-gp induction by rifampin was negligible in MDCK-cells while MDCK cells appeared to be feasible for P-gp inhibition studies. Taken all together, the present study suggests that Caco-2 cells might be more applicable to the BCS drug classification than MDCK-cells, although MDCK cells may provide some advantage in terms of capacity and speed in early ADME screening process.

The Effects of MDCK Cell on Low Dose Irradiation (저선량 방사선 조사가 배양 세포에 미치는 효과)

  • Lee, Song-Jae;Chang, Jae-Chul
    • The Journal of Korean Society for Radiation Therapy
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    • v.9 no.1
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    • pp.106-112
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    • 1997
  • The present study was undertaken to investigate the effects of cultured MDCK cell line on the cell viability and the activities of superoxide dismutase(SOD). catalase, change of FOX 1 with low dose radiation. When MDCK cells were irradiated low dose (less than 50 cGy), the cell viability remains high after 2 hrs, but few changes after 24 hrs. In the low dose irradiated MDCK cells, the activities of SOD and catalase were increased with compared to control group and high dose. But the content of $H_2O_2$(FOX 1) was decreased. These results suggest that the cultured MDCK cells probably were induced expression of defense mechanism.

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Changes of Glutathione S-transferase Activity in MDCK Cells with Influenza Virus Type A (Influenza Virus Type A 감염 MDCK세포에 있어서 Glutathione S-transferase의 활성 변동)

  • 김병렬;박남표;윤종국;전태원;이상일
    • Biomedical Science Letters
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    • v.6 no.1
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    • pp.19-28
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    • 2000
  • This study was performed to evaluate the activities of glutathione S-transferase (GST) and lactate dehydrogenase (LDH) in Maddin-Darby canine kidney (MDCK) cells infected with virus and/or treated with amantadine. On cell morphological findings, monolayer fractions in MDCK cells infected with virus were exfolated more than 80% in 1 TCID$_{50}$ group and that in 10 TCID$_{50}$ were completely exfolated after 3 days during infectious process. In proportion to the dose of amantadine, activities of GST and LDH of MDCK cells were significantly decreased and those of LDH in medium fraction were more significantly increased compared with control. According to in both dose and time of virus innoculation, activities of GST and LDH in MDCK cells were significantly decreased in 1 and 10 TCID$_{50}$ infected cells after 3 days. LDH activities in infectious medium were remarkably rised at 10 fold. In case of the cell line inoculated with type A 100 TCID$_{50}$ and additionally treated with amantadine, the decreasing rate to the control in activities of GST and LDH was lower than that in those in case of that infected with virus only. These results suggested that virus infection and amantadine treatment may effect the activity of the detoxicating enzyme in the target cells.

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Effect of WHW, a polyherbal medicine for the treatment of chronic renal failure on staurosporin-induced apotosis in MDCK cells (만성신부전 한약제제 WHW의 신장세포에서의 Staurosporine 유도 세포사멸에 대한 억제 효과)

  • Bae, Hyo-Sang;Yoon, Cheol-Ho;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.27 no.4
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    • pp.45-51
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    • 2012
  • Objectives : WHW is a polyherbal medicine for the treatment of chronic renal failure (CRF). WHW previously reported various biological property such as anti-inflammation, anti-oxidation and anti-renal fibrosis in CRF. This study aimed to investigate the anti-apoptotic effect of WHW on staurosporin(SSP)-induced apoptosis in canine kidney epithelial cells (MDCK). Methods : MDCK cells were treated with different concentrations of WHW (0.1, 0.2, 0.5 and $1mg/m{\ell}$) for 1 h, and then induced apoptosis by treatment of SSP ($1{\mu}M$) for 24 h. Cell viability was measured by WST-1 assay. The expression of apoptotic proteins such as caspase-3, Bax and Bcl-2 was determined by Western blot. Caspase-3 activity and ROS levels were also measured by their commercial available assay kits. Cell apoptosis was observed by Hoechst and DNA fragmentation. Results : WHW significantly increased the cell viability on SSP-treated MDCK cells. WHW inhibited SSP-induced expression of apoptotic proteins such as caspase-3 and Bax, and significantly decreased caspase-3 activity in MDCK cells. WHW significantly decreased SSP-induced production of ROS, and suppressed SSP-induced chromatin condensation and DNA fragmentation in MDCK cells. Conclusions : These results suggest that WHW has an anti-apoptotic effect in renal cells through suppressing the expression of apoptotic proteins, ROS production and DNA damages.

Generation of a High-Growth Influenza Vaccine Strain in MDCK Cells for Vaccine Preparedness

  • Kim, Eun-Ha;Kwon, Hyeok-Il;Park, Su-Jin;Kim, Young-Il;Si, Young-Jae;Lee, In-Won;Kim, Se mi;Kim, Soo-In;Ahn, Dong-Ho;Choi, Young-Ki
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.997-1006
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    • 2018
  • As shown during the 2009 pandemic H1N1 (A(H1N1)pdm09) outbreak, egg-based influenza vaccine production technology is insufficient to meet global demands during an influenza pandemic. Therefore, there is a need to adapt cell culture-derived vaccine technology using suspended cell lines for more rapid and larger-scale vaccine production. In this study, we attempted to generate a high-growth influenza vaccine strain in MDCK cells using an A/Puerto/8/1934 (H1N1) vaccine seed strain. Following 48 serial passages with four rounds of virus plaque purification in MDCK cells, we were able to select several MDCK-adapted plaques that could grow over $10^8PFU/ml$. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells ($PB1_{D153N}$, $M1_{A137T}$, and $NS1_{N176S}$). In addition, we confirmed that in the context of the high-growth A/PR/8/34 backbone, A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), and A/environment/Korea/deltaW150/2006 (H5N1) also showed significantly enhanced growth properties (more than $10^7PFU/ml$) in both attached- and suspended-MDCK cells compared with each representative virus and the original PR8 vaccine strain. Taken together, this study demonstrates the feasibility of a cell culture-derived approach to produce seed viruses for influenza vaccines that are cheap and can be grown promptly and vigorously as a substitute for egg-based vaccines. Thus, our results suggest that MDCK cell-based vaccine production is a feasible option for producing large-scale vaccines in case of pandemic outbreaks.

Mercuric Chloride Induces Apoptosis in MDCK Cells (Mercuric Chloride에 의한 MDCK 세포의 세포사멸)

  • Lee, Ju-Hyoung;Youm, Jung-Ho;Kwon, Keun-Sang
    • Journal of Preventive Medicine and Public Health
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    • v.39 no.3
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    • pp.199-204
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    • 2006
  • Objectives: Mercury is a hazardous organ-specific environmental contaminant. It exists in a wide variety of physical and chemical states, each of which has unique characteristics for the target organ specificity. Exposure to mercury vapor and to organic mercury compounds specifically affects the CNS, while the kidney is the target organ for inorganic Hg compounds. Methods: In this study, mercury chloride $(HgCl_2)$ was studied in a renal derived cell system, i.e., the tubular epithelial Madin-Darby canine kidney (MDCK) cell line, which has specific sensitivity to the toxic effect of mercury. MDCK cells were cultured for 6-24 hr in vitro in various concentrations (0.1-100 M) of $HgCl_2$, and the markers of apoptosis or cell death were assayed, including DNA fragmentation, caspase-3 activity andwestern blotting of cytochrome c. The influence of the metal on cell proliferation and viability were evaluated by the conventional MTT test. Results: The cell viability was decreased in a time and concentration dependent fashion: decreases were noted at 6, 12 and 24 hr after $HgCl_2$, exposure. The increases of DNA fragmentation were also observed in the concentrations from 0.1 to 10 M of $HgCl_2$ at 6 hr after exposure. However, we could not observe DNA fragmentation in the concentrations more than 25 M because the cells rapidly proceeded to necrotic cell death. The activation of caspase-3 was also observed at 6 hr exposure in the $HgCl_2$ concentrations from 0.1 to 10 M. The release of cytochrome c from the mitochondria into the cytosol, which is an initiator of the activation of the caspase cascade, was also observed in the $HgCl_2-treated$ MDCK cells. Conclusions: These results suggest that the activation of caspase-3 was involved in $HgCl_2-induced$ apoptosis. The release of cytochrome c from the mitochondria into the cytosol was also observed in the $HgCl_2-treated$ MDCK cells. These findings indicate that in MDCK cells, $HgCl_2$ is a potent inducer of apoptosis via cytochrome c release from the mitochondria.

Comparative susceptibility of different cell lines for culture of Toxoplasma gondii in vitro (톡소플라스마 곤디의 세포내 배양에 있어서 세포 주에 따른 감수성 비교)

  • 박병규;문형로
    • Parasites, Hosts and Diseases
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    • v.31 no.3
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    • pp.215-222
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    • 1993
  • In order to establish a useful cell culture system for T gondii we compared the degree of proliferation of T gondii tachyzoites among 8 different cell lines: 2 kinds of normal animal cells (MDCK-canine kidney cells; Vero-monkey kidney cells) and 6 kinds of human tumor cells (A 549, PC 14-lung cancer cells; SNU 1, SNU 16. Mlm 45-stomach cancer cells; HL-60-promyelocytic leukemia cells), through morphological observation and 3H-uracil uptake assay. The degree of susceptibility to infection with T gondii tachyzoites was highest in A 549 and PC 14 cells, medium in Vero, HL-60, MDCK and SNU 1, and lowest in SNU 16 and MBm 45 cells. The kinetics of T gondii multiplication during the post-Infection 60 hours were higllly dependent upon the dose of tachyzoites administered and the duration among the 8 tested fur the growth and multiplication of T gondii in vitro.

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Effect of Chrysanthemum Morifolium Extracts on the Synthesis of Laminin of Madin-Darby Canine Kidney Cells (감국(甘菊)이 MDCK 세포의 Laminin 합성에 미치는 영향)

  • Na, Ho-Jeong;Jeon, So-Ra;Cha, Dong-Seok;Eun, Jae-Soon;Lim, Jong-Pil;Shin, Tae-Yong;Oh, Chan-Ho;Yang, Jae-Heon;Kim, Dae-Keun;Leem, Jae-Yoon;Chae, Byeong-Suk;Kim, Sung-Zoo;Jung, Yen-Ok;Jeong, Won-Hwan;Jeon, Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.3
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    • pp.709-713
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    • 2007
  • Basement membranes (BMs) are extracellular matrices associated with epithelia, endothelia, muscle, fat and peripheral nerve. They are involved in cell survival, migration, differentiation. BMs functions also include tissue formation and provide mechanical stability as a selective barriers. Laminins are heterotrimeric glycoproteins found in BMs and have a crucial role in cell adhesion and signalling. Madin-Darby canine kidney (MDCK) cells are the best established mammalian model for studying epithelial cell biology The cells form an epithelial monolayer, with tight junctions separating an apical surface from a basolateral membrane facing the filter support and neighboring cells. In this study, using MDCK cells, the synthesis of the BM protein such as laminin with or without methanol extract of Chrysanthemum morifolium (CM) stimulation was analyzed by immunoblotting and CM showed significant increased cell density and enhanced synthesis of laminin.

Development and Application of Non-Destructive-Type Device of Ingredients in Mulberry Leaf Tea (뽕잎 차에서의 성분 비파괴형 기기 기술 개발 및 응용)

  • Cheun, Byeung-Soo;Min, Je-Ho
    • The Journal of the Korea institute of electronic communication sciences
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    • v.8 no.10
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    • pp.1595-1600
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    • 2013
  • In the present study,effect of natural mulbelly leaf tea on DMEM MDCK cell line. Mulbelly leaf teas inhibited the proliferation in primery MDCK cells in a dose dependent manner. These results show that mulbelly teas potent inhibits the reactive oxygen species (ROS) and not destruction a component. Therefore, mulbelly teas might improve overall quality of the color and taste. And might be applied newly to development of componant mulbelly teas quality and biochemical change by ROS in living things.

The effect of microfilament inhibitor on the Cryptosporidium infection in vitro

  • Yu, Jae-Ran;Choi, Saung-Don
    • Parasites, Hosts and Diseases
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    • v.38 no.4
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    • pp.257-261
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    • 2000
  • This study was focused on the effects of microfilament inhibitor, Cytochalasin D (CD) on the invasiveness of sporozoites of Cryptosporidiun spp. into the host cells. MDCK and AGS cell lines were used as host cells for C. parvum and C. muris, respectively. When MDCK cells were pretreated with CD for 1 hr before inoculation of the sporozoites, C. parvum infection was significantly inhibited when compared to the control cells. These inhibitory effects of CD on the rate of infection were dose-dependent. In addition, C. muris infection was hampered when AGS cell lines were pretreated with CD. However, the capability of invasiveness of the sporozoites into the host cells was not greatly influenced by the pretreatment of sporozoites with CD before infection. These results suggest that microfilaments of host cells, rather than parasites, play an important role for the invasion of Cryptosporidium spp.

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