The objective of this study was to improve the cell-substrate interactions between skin cells and a biodegradable elastomeric matrix, part of a cell-seeded skin substitute for the treatment of large-scale deep dermal skin defects (i.e. burn wounds). Polyactive, a synthetic biodegradable elastomeric copolymer, was used as the constituent of the bilayered matrix. It consists of a dense toplayer seeded with epidermal keratinocytes and a macroporus underlayer, which may be seeded with dermal fibroblasts. Although former studies demonstrated the suitability of the copolymer as a substrate for these skin-derived cell types, we aimed to improve the bilayered matrix' seeding efficiency. Using radio frequency glow discharge (RFGD) pretreatment significantly improved the adherence and growth of SVK14 epithelial cells seeded on the dense copolymeric toplayers and on non-tissue grade plastics, approximating tissue culture polystyrene values. With scanning electron microscopy (SEM), early epithelial cell-substrate interactions were investigated. Seeding efficiency and growth of dermal fibroblasts into the porous underlayers was improved as was visualized with the SEM and confocal scanning laser microscopy. It is concluded that RFGD pretreatment is a cost-effective measurement for improving cell-substrate properties of the investigated copolymers.