• Journal of Pharmaceutical Investigation
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• 제38권3호
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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.

• 대한방사선치료학회지
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• 제9권1호
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• pp.106-112
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• 1997

MDCK 세포를 한국 세포주 은행으로부터 분주 받아 배양한 후 저선량 gamma선 조사에 대한 세포수준에서 방사선 효과를 알아보기 위하여 세포 증식능의 변화, Superoxide dismutase(SOD)와 catalase, FOX I의 함량 변화를 측정 검토하였다. 그결과, 저선량의 방사선를 조사했을 때, 세포의 증식능은 방사선 조사후 2시간에서 10 cGy는 $95.1\%$, 50cGy는 $96.4\%$로 대조군보다 약간의 감소가 나타났으나, 24시간 후에서 10cGy는 $96.7\%$, 50cGy는 $73.1\%$로 선량이 증가함에 따라 세포 증식능은 감소하였다. 저선량 조사에 의한 MDCK 세포의 SOD 활성도는 전반적으로 증가하였고, Mn-SOD 활성 역시 증가하였다. 세포 내의 $H_{2}O_2$의 양을 측정한 FOX I에서 선량이 증가함에 따라 감소하였으며 catalase 효소의 함량은 대조군보다 증가되는 경향을 보였다. 이와 같은 결과로 볼 때 저선량 방사선 조사에 대한 효과는 세포내부의 자체적인 방어기작의 발현으로 인한 결과라고 생각된다.

• 대한의생명과학회지
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• 제6권1호
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• pp.19-28
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• 2000

농도와 기간에 따른 바이러스 감염 및 항바이러스제인 amantadine병행 첨가 시 Maddin Darby canine kidney (MDCK) 세포 내의 free radical 해독계 효소인 glutathione S-transferase (GST)와 lactate dehydrogenase (LDH)의 활성 변동을 상호비교 관찰하였다. 바이러스에 감염된 MDCK세포는 감염 3일 후 1 TCID$_{50}$ 군은 80%이상, 10 TCID$_{50}$ 군은 거의 대부분의 단층세포가 탈락되는 병변 효과가 나타났다. Amantadine cytotoxic dose %가 증가함에 따라 MDCK 세포 내의 GST 및 LDH 활성은 농도에 따라 유의하게 감소되었고, 감염배지 내 LDH 활성은 대조군 보다 유의한 증가를 보였다. 인플루엔자 바이러스 type A 접종농도와 기간에 따른 MDCK세포 내의 GST및 LDH활성은 1 및 10 TCID$_{50}$ 감염군에서 감염 3일 후부터 유의하게 감소되었고, 감염배지 내의 LDH활성은 10배 이상 증가되었다. 인플루엔자 바이러스 type A 100 TCID$_{50}$ 감염과 amantadine병행 첨가 시 GST 및 LDH 활성은 바이러스만 감염한 군 보다 MDCK세포 내에서 대조군에 비하여 감소율이 낮았고, 감염배지 중 LDH 활성 역시 증가율이 낮았다. 또한 바이러스 감염 후 amantadine 90 $\mu\textrm{g}$/ml 첨가 시에 세포 내와 감염배지 중에서 가장 낮은 감소와 증가를 나타냈다.

• 대한본초학회지
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• 제27권4호
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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.

• Journal of Microbiology and Biotechnology
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• 제28권6호
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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.

• Journal of Preventive Medicine and Public Health
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• 제39권3호
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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.

• Parasites, Hosts and Diseases
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• 제31권3호
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• pp.215-222
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• 1993

Toxoplasmngon gondii의 세포내 배양에 적합한 숙주 세포 주를 찾기 위하여 정상 세포 2종류(MDCK-canine kidney cells; Vero-monkey kidney cells) 및 암세포 6종류(A 549, PC 14-human lung cancer cells; SNU 1, SNU 16, MKN 45-human stomach cancer cells; HL-60-human promyelocytic leukemia cells)를 대상으로 하여 각 세포 주의 T.gondii 감염에 대한 감수성을 형태학적 관찰 및 3H-uracil 흡수 시험을 통하여 비교하였다. T.gondii 대한 감수성은 A 549 및 PC 14 세포가 가장 높았고, Vero, HL-60, MDCK 및 SNU 1 세포가 그 다음, SNU 16 및 MKN 45 세포는 가장 감수성이 낮았다. 또한 각 세포 주에 있어서 T.gondii 감염 후 충체증식 정도를 정량화하여 12시간, 36시간 및 60시간에 각각 측정한 바 충체 수를 적게($2{\times}10^5/ml$) 투여했을 때는 A 549, PC 14, Vero, MDCK 세포들에서 감염 60시간까지 충체의 분열 증식이 계속 증가하였고, 충체 수를 많이($50{\times}10^5/ml$) 주입하였을 때는 대부분의 세포들에서 감염 12시간에 최고의 증식을 보이다가 이후 증식이 감소하였다. 이상의 결과로 보아 기son사거 분리 계대 및 충주(strain) 확립을 위해서는 A 549 및 PC 14 세포가 가장 적합할 것으로 판단되며, 충체 주입 수 및 배양 시간별로 충체의 증식 정도가 다름을 알 수 있었다.

• 동의생리병리학회지
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• 제21권3호
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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.

• Journal of Microbiology and Biotechnology
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• 제33권12호
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• pp.1576-1586
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• 2023

Vaccination is the most effective method for preventing the spread of the influenza virus. Cell-based influenza vaccines have been developed to overcome the disadvantages of egg-based vaccines and their production efficiency has been previously discussed. In this study, we investigated whether treatment with forskolin (FSK), an adenylyl cyclase activator, affected the output of a cell-based influenza vaccine. We found that FSK increased the propagation of three influenza virus subtypes (A/H1N1/California/4/09, A/H3N2/Mississippi/1/85, and B/Shandong/7/97) in Madin-Darby canine kidney (MDCK) cells. Interestingly, FSK suppressed the growth of MDCK cells. This effect could be a result of protein kinase A (PKA)-Src axis activation, which downregulates extracellular signal-regulated kinase (ERK)1/2 activity and delays cell cycle progression from G1 to S. This delay in cell growth might benefit the binding and entry of the influenza virus in the early stages of viral replication. In contrast, FSK dramatically upregulated ERK1/2 activity via the cAMP-PKA-Raf-1 axis at a late stage of viral replication. Thus, increased ERK1/2 activity might contribute to increased viral ribonucleoprotein export and influenza virus propagation. The increase in viral titer induced by FSK could be explained by the action of cAMP in assisting the entry and binding of the influenza virus. Therefore, FSK addition to cell culture systems could help increase the production efficiency of cell-based vaccines against the influenza virus.

• 한국전자통신학회논문지
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• 제8권10호
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• pp.1595-1600
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• 2013

기존의 기기분석에 있어서의 미비한 점을 보완한 물질 분석 과정에서 주요 성분의 파괴를 최소화하고져 비파괴형 원적외선 도자기 가마를 이용하여 비파괴형 성분물질을 유지할 수 있는 기기 분석 장치를 개발 하였다. 또한 광폭증폭관(PMT) 기기를 사용하여 뽕잎 차에 함유된 항산화 물질의 형태적 효과를 기존의 기기분석기와 원적외선 도자기 기기를 사용한 것과의 차별화 실험을 통해 그 효과를 측정 하였다. 그 결과 특히 과산화수소를 이용한 항산화 효과를 측정한 결과 성분이 파괴되지 않음을 측정하여 그 효과가 인정 되었기에 이를 보고한다.