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p38 signaling and receptor recycling events in a microfluidic endothelial cell adhesion assay.

PloS one (2013-06-14)
Dwayne A L Vickers, Emma J Chory, Megan C Harless, Shashi K Murthy
RESUMEN

Adhesion-based microfluidic cell separation has proven to be very useful in applications ranging from cancer diagnostics to tissue engineering. This process involves functionalizing microchannel surfaces with a capture molecule. High specificity and purity capture can be achieved using this method. Despite these advances, little is known about the mechanisms that govern cell capture within these devices and their relationships to basic process parameters such as fluid shear stress and the presence of soluble factors. This work examines how the adhesion of human endothelial cells (ECs) is influenced by a soluble tetrapeptide, Arg-Glu-Asp-Val (REDV) and fluidic shear stress. The ability of these ECs to bind within microchannels coated with REDV is shown to be governed by shear- and soluble-factor mediated changes in p38 mitogen-activated protein kinase expression together with recycling of adhesion receptors from the endosome.

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Sigma-Aldrich
(3-Mercaptopropyl)trimethoxysilane, 95%
Sigma-Aldrich
Milli-Mark® Anti-Integrin β1-FITC Antibody, activated, clone HUTS-4, clone HUTS-4, Milli-Mark®, from mouse