Source:http://linkedlifedata.com/resource/pubmed/id/16962665
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
2007-3-12
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pubmed:abstractText |
Vascular permeability is a pathologic process in many disease states ranging from metastatic progression of malignancies to ischemia-reperfusion injury. In order to more precisely study tissue, and more specifically cell layer permeability, our goal was to create a fluorescence-based assay which could quantify permeability without radioactivity or electrical impedance measurements. Human aortic endothelial cells were grown in monolayer culture on Costar-Transwell clear polyester membrane 6-well cell culture inserts. After monolayer integrity was confirmed, vascular endothelial growth factor (VEGF(165)) at varying concentrations with a fixed concentration of yellow-green fluorescent 0.04 microm carboxylate-modified FluoSpheres microspheres were placed in the luminal chamber and incubated for 24 h. When stimulated with VEGF(165) at 20, 40, 80, and 100 ng/ml, this assay system was able to detect increases in trans-layer flux of 8.2+/-2.4%, 16.0+/-3.7%, 41.5+/-4.9%, and 58.6+/-10.1% for each concentration, respectively. This represents the first fluorescence-based permeability assay with the sensitivity to detect changes in the permeability of a cell layer to fluid flux independent of protein flux; as well as being simpler and safer than previous radioactive-and impedance-based permeability assays. With the application of this in vitro assay to a variety of pathologic conditions, both the dynamics and physiology relating to cellular permeability can be more fully investigated.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Fluorescent Dyes,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/VEGFA protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Vascular Endothelial Growth Factor A
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pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
0165-022X
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
10
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pubmed:volume |
70
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
329-33
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pubmed:meshHeading |
pubmed-meshheading:16962665-Biophysics,
pubmed-meshheading:16962665-Capillary Permeability,
pubmed-meshheading:16962665-Cell Membrane Permeability,
pubmed-meshheading:16962665-Cells, Cultured,
pubmed-meshheading:16962665-Endothelial Cells,
pubmed-meshheading:16962665-Fluorescent Dyes,
pubmed-meshheading:16962665-Humans,
pubmed-meshheading:16962665-Microspheres,
pubmed-meshheading:16962665-Models, Biological,
pubmed-meshheading:16962665-Recombinant Proteins,
pubmed-meshheading:16962665-Vascular Endothelial Growth Factor A
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pubmed:year |
2007
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pubmed:articleTitle |
A novel fluorescence-based cellular permeability assay.
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pubmed:affiliation |
Section of Vascular and Endovascular Surgery, Department of Surgery, Univeristy of California, San Diego Medical Center, 200 West Arbor Drive, San Diego, CA 92103-8402, USA.
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pubmed:publicationType |
Journal Article
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