Abstract
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.
Highlights
Endothelial cells (ECs) line the blood vessel walls and serve as an interface for blood flow [1,2]
These changes were assessed by first incubating human umbilical vein endothelial cells (HUVECs) in the mentioned REDV concentrations to block all REDV binding receptors followed by flow within REDV functionalized microfluidic channels to evaluate cell adhesion
The adhesion aspect of this data shows that incubation in soluble REDV followed by flow of HUVECs through REDV-coated microchannels for,45 s residence time causes a decrease in cell adhesion within the microchannels for concentrations of 50 mg/mL soluble REDV and below
Summary
Endothelial cells (ECs) line the blood vessel walls and serve as an interface for blood flow [1,2]. The hemodynamic force from the luminal blood, together with adhesive forces between cell surface anchoring proteins (integrins) and the basement membrane (basal lamina) contribute to a complex set of mechanical signals that are known to regulate vascular function through multiple, mechanotransduction-related signaling pathways [3,4] Based on their important role in the cardiovascular system attention has been focussed on isolating both immature and mature ECs from heterogeneous starting material for applications such as tissue engineering [5,6,7]. Adhesion-based cell separation has proven to be very useful in a wide range of applications, ranging from cancer diagnostics to tissue engineering because it eliminates the need for sample pre-processing to bind fluorescent or magnetic tags [8,9,10,11] This approach involves functionalizing microfluidic channels with molecules that bind to one or more cell types that are captured from a flowing stream. This assumption enables the design of microfluidic capture systems based on simple profiles of cell adhesion as function of shear stress [9]
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