Fabrication of substrates with curvature for cell cultivation by alpha-particle irradiation and chemical etching of PADC films

In the present paper, we developed a microfabrication technology to generate cell-culture substrates with identical chemistry and well-defined curvature. Micrometer-sized pits with curved surfaces were created on a two-dimensional surface of a polymer known as polyallyldiglycol carbonate (PADC). A PADC film was first irradiated by alpha particles and then chemically etched under specific conditions to generate pits with well-defined curvature at the incident positions of the alpha particles. The surface with these pits was employed as a model system for studying the effects of substrate curvature on cell behavior. As an application, the present work studied mechanosensing of substrate curvature by epithelial cells (HeLa cells) through regulation of microtubule (MT) dynamics. We used end-binding protein 3-green fluorescent protein (EB3-GFP) as a marker of MT growth to show that epithelial cells having migrated into the pits with curved surfaces had significantly smaller MT growth speeds than those having stayed on flat surfaces without the pits.