development of the evaluation system for barrier functions of engineered epithelial lumens

Abstract

We have investigated the effects of a diameter of engineered epithelial lumen on cellar architectures and a barrier function. For this investigation, we have developed a system to evaluate the barrier function of engineered epithelial lumens. To test the utility of our system, we constructed the engineered epithelial lumens by culturing Madin–Darby Canine Kidney cells (MDCK) on the gold wires with different diameters ranging from 50 μm–200 μm. Confocal laser scanning microscopy revealed that long actin stress fibers and a low focal adhesion density were observed at the gold wire diameter of 200 μm, whereas the mesh-like morphology consisted of short actin stress fibers and high focal adhesion densities were found at the gold wire diameters of 50 μm and 100 μm. The expression pattern of ZO-1 that localizes at the tight junction was independent on the gold wire diameter. The electrical impedance measurement indicates that the barrier function for the samples constructed at the gold wire diameter of 200 μm was significantly higher than those at the gold wire diameters of 50 μm and 100 μm. The difference in the barrier functions of epithelial lumens might be attributed to the changes in cellular architectures with increasing the curvature of gold wire.