タイトジャンクションのバリア機能評価用マイクロデバイス

抄録

We aim to develop a microdevice that measures material permeation via epithelial tight junction (TJ) to evaluate its barrier function. A microdevice is equipped with square pillar microchamber filled with collagen gel. Thus, when epithelial cells are cultured on its surface, cells do not invade into the microchamber, so that a flat epithelial sheet is formed. If the inner space of the microchamber is closed by a cell sheet, the inflow and outflow of the matter is restricted. If the permeability of TJ is increased by drugs, the target substance concentration in the microchamber changes due to the diffusion. In this study, the mass transfer of fluorescent dextran, a model substance, between the chamber and the bulk space was measured, in correlation to the morphology of MDCK cells on the microchamber device, using a confocal laser scanning microscope (CLSM). On the first day of cell culture, it was confirmed that the cells did not enter the microchamber and adhered on the collagen gel, and the leakage of fluorescent dextran was suppressed. However, after the second day of cell culture, cells invaded into collagen gel. The development of TJ requires about 3 days of culture, so we will study the concentration and hardness of collagen gel to develop a device that can measure substance permeation under long-term culture conditions.