Furusawa Kazuya, Mizutani Takeomi, Sasaki Naoki
Faculty of Advanced Life Science, Hokkaido University, Kita-ku, Kita 10 Nishi 8, Sapporo, Hokkaido, Japan.
Regen Ther. 2016 Mar 22;3:82-89. doi: 10.1016/j.reth.2016.02.010. eCollection 2016 Mar.
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.
我们研究了工程化上皮管腔直径对细胞结构和屏障功能的影响。为此项研究，我们开发了一种评估工程化上皮管腔屏障功能的系统。为测试我们系统的实用性，我们通过在直径范围为50μm至200μm的金线表面培养犬肾上皮细胞（MDCK）构建工程化上皮管腔。共聚焦激光扫描显微镜显示，在200μm金线直径处观察到长肌动蛋白应力纤维和低粘着斑密度，而在50μm和100μm金线直径处发现由短肌动蛋白应力纤维和高粘着斑密度组成的网状形态。定位在紧密连接处的闭合蛋白1（ZO-1）的表达模式与金线直径无关。电阻抗测量表明，在200μm金线直径处构建的样品的屏障功能显著高于在50μm和100μm金线直径处构建的样品。上皮管腔屏障功能的差异可能归因于随着金线曲率增加细胞结构的变化。
