A microfluidic chip for permeability assays of endothelial monolayer

Abstract

Endothelial cell monolayer (EM), acting as a barrier between blood and tissue, plays an important role in pathophysiological processes. Here we describe a novel microfluidic chip that is applied for convenient and high throughput in vitro permeability assays of EM. The chip included a gradient generator and an array of cell culture chambers. A microporous membrane as a scaffold component was built between a polydimethylsiloxane (PDMS) layer and a glass substrate to grow EM. Cell culture chambers were separated by microchannels and microvalves. The concentration gradient of compound solutions could be generated automatically and affected EM in different chambers. The permeability of EM at different time with histamine stimulation was in situ measured by the fluorescence detection of the leaked tracer. The existence of continuous flow in the channels allowed EM in a dynamic microenvironment and increased the amount of tracer through the EM, comparing to transwell assays. According to the prototype chip, the chip with a bigger array of cell culture chambers could be achieved easily and applied in the high throughput screening for drugs.

Novel Stem Cell–Based Drug Discovery Platforms for Cardiovascular Disease

“…116,117 These assays are highly quantitative and are able to produce time-dependent permeability data able to discern transient effects. Miniaturization and parallelization of these assays, both fluorescent-based macromolecule transport 118 and commercially available 96-well platforms and culture plates for measuring transendothelial electrical resistance, permit screening assays to use endothelial permeability as a direct functional output.…”

A Membrane-integrated Microfluidic Device to Study Permeation of Nanoparticles through Straight Micropores toward Rational Design of Nanomedicines

“…9,10 The size of a microfluidic channel can be adjusted to that of a microvessel, and various vascular cells can be cultured in the channel under blood flow-like flow conditions. [11][12][13] In one approach, a porous membrane was integrated into a microfluidic device and utilized to evaluate the permeability of an endothelial monolayer on the membrane against fluorescein isothiocyanatelabeled bovine serum albumin (FITC-BSA) [14][15][16] and lipid-coated nanoparticles. 16 However, the pores within the monolayer are irregular in size and shape, as shown in previous studies.…”

A high-throughput permeability assay platform for shear stress characterization of endothelial cells

“…Microfluidic in vitro models, which are advantageous over in vivo models because they can generate well-defined shear stress in a controllable and repeatable manner, have been used to study shear stress effects on morphology [4] and permeability [5], but no previous system has managed to simultaneously induce the full-spectrum range of shear stress and investigate effects on permeability. Microfluidic systems have induced a shear stress range with geometric variation [6], but the non-uniform shear stress applied to the cell populations make such channels unfeasible for nonimaging measurement methods.…”