Journal
LAB ON A CHIP
Volume 11, Issue 24, Pages 4235-4240Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c1lc20722a
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Funding
- European Commission [CP-FP 214566-2, IRSES-GA-2009-247641]
- National Basic Research Program of China (973 Program) [2007CB714507, 2011CB933600]
- Science Fund for Creative Research Groups [20921062]
- National Natural Science Foundation of China [20875072]
- Chinese Scholar Council
- Grants-in-Aid for Scientific Research [22350104] Funding Source: KAKEN
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We present a method that allows patterning cells and shear flow conditions for endothelial cell based assays. This method is novel in combining (1) cell culture on the surface of a substrate both topographically and chemically patterned; (2) multi-shear flow assays after covering the cell substrate with a microfluidic cover plate containing microchannels of different channel widths, and (3) conventional immunostaining assays after removal of the cover plate. This method has the advantage of performing cell cultures and immunoassays in standard cell biology environments with open access, facilitating the formation of confluent cell layers and the observation of cell responses to shear-flow and drug stimulations. To obtain multi-shear stress conditions, a single channel with stepwise increasing channel widths was patterned on the surfaces of both the substrate and the microfluidic cover plate. As results, we observed excellent viability of endothelial cells in the whole range of applied shear stresses (0-25 dyn cm(-2)) and shear stress dependent cytoskeleton remoulding, activation of von Willebrand factor (vWF), and re-organisation of angiogenesis factors such as tetra peptide acetyl-Ser-Asp-Lys-Pro (AcSDKP) of endothelial cells. To validate this approach for drug analysis, we also studied drug effects under shear stress conditions. Our results indicate that the drug effect of combretastatin A-4, an antitumour vascular targeting drug, could be significantly enhanced under shear flow conditions.
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