Convenient fabrication of three-dimensional cell-culture substrates through introduction of micrometer-size pores on polyallyldiglycol carbonate polymer films

We explored the fabrication of three-dimensional (3D) substrates by creating micrometer-size pores on polyallyldiglycol carbonate (or PADC) polymer films through irradiation of the film by alpha particles and subsequent chemical etching. HeLa cells cultured on these 3D substrates were observed using scanning electron microscope. Multiple directions and multiple layers of HeLa cells were observed to have grown in the pores, with normal nuclei and cell membranes as well as good cell spreading. For the cells cultured in 3D substrates with or without additional small pores, no significant differences were observed between their vinculin expression profiles, which were in contrast to the observation made for cells cultured on 2D substrates showing that small pores could enhance vinculin expression. The presence of the large pores and/or the enhanced biocompatibility of the substrate in the present experiments might be the reasons. The protrusions of cells were confined by the small pores, which was similar to the observation made for cells cultured on 2D substrates.