title: Patterning cells and shear flow conditions: convenient observation of endothelial cell remoulding, enhanced production of angiogenesis factors and drug response.
abstract: 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 anti-tumour vascular targeting drug, could be significantly enhanced under shear flow conditions.
authors: Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences and State Key Laboratory of Virology, Wuhan University, Wuhan 430072, P. R. China.
authors: Li
authors: L
authors: Wang
authors: Zhi-Ling
authors: ZL
authors: Zhang
authors: Joanna
authors: J
authors: Wdzieczak-Bakala
authors: Dai-Wen
authors: DW
authors: Pang
authors: Jianmiao
authors: J
authors: Liu
authors: Yong
authors: Y
authors: Chen
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000378:metabolism*; D002478:Cells, Cultured; D003599:Cytoskeleton; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects*; D015200:Fluoroimmunoassay; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D009842:Oligopeptides / Q000378:metabolism*; D033081:Shear Strength; D013267:Stilbenes / Q000494:pharmacology*; D014841:von Willebrand Factor / Q000378:metabolism
chemical_list: D043925:Angiogenesis Inducing Agents; D009842:Oligopeptides; D013267:Stilbenes; D014841:von Willebrand Factor; C058504:goralatide; C058728:fosbretabulin