A polyimide based on alicyclic units, such as epiclon (5-(2,5-dioxotetrahydrofurfuryl)-3-methyl-3-cyclohexene-1,2-dicarboxylic acid anhydride), was investigated from the point of view of its surface features to check their suitability for cell culture applications. Atomic force microscopy data revealed fractal and isotropic surfaces with nanoscale roughness and peaks placed at much smaller distances comparing to the cell size scale, favoring a good compatibility of the synthesized material with the biological medium, particularly after plasma treatment. Surface tension properties were determined in order to evaluate the interactions at the bio-interface affecting the adherence behavior of cell-binding proteins to the sample surface. In vitro experiments regarding the cytocompatibility and population tendency reveal that polyimide allows cells to adhere and to proliferate onto the surface. These tests indicate that the studied epiclon-based polyimide is not cytotoxic and can be recommended as good candidate for cell culture substrate in tissue engineering, especially after plasma treatment.