Establishment of cell patterning method utilising nanopore structure with cell non-adhesive effect

Abstract

This paper reports on a novel cell patterning method that uses nanoscale porous structures called nanopores, which are expected to prevent cell adhesion. The adhesion prevention area can be formed selectively on the surface of the culture substrate in combination with photolithography. In this study, a culture substrate having both a cell non-adhesion region and a cell adhesion region on the same surface was fabricated. The cells were seeded on the culture substrate, and their forms in both regions were compared and discussed. The result shows that the number of cells extending the pseudopodium in the cell non-adhesion region was 2.6% of that in the cell adhesion region. This demonstrates that the effect of cell adhesion prevention is determined only by the characteristics of the uneven nanoscale structure. The merit of this method is its effectiveness, avoiding the necessity of applying light irradiation or an electric signal to a cell. In addition, even when many chemical treatments are required before and after culturing, effects such as reduction of cost and time, and avoidance of interference between multiple treatments can be expected. Furthermore, in anticipation of being linked to the technology of producing cell sheets composed of cells of arbitrary shapes and multiple types, a method has been established that can form nanopores that effectively prevent cell adhesion on a large area at low cost. A culture substrate for arbitrary cell patterns can be easily prepared using general photolithography in a 5 min dry etching process.