Electrospun polymeric micro- and nanofibers offer the opportunity to reproduce the filamentous structure of the extracellular matrix in native tissues. The ability to electrospin extracellular matrix proteins offers the additional benefit of being able to create a chemical and structural microenvironment for cells in order to influence their behavior during scaffold seeding and tissue formation. This offers significant advantages over unspecific protein adsorption or functionalisation with adhesion promoting peptide sequences. However, producing electrospun fibers from pure protein solutions remains difficult and costly. A more accessible approach is to introduce proteins into electrospun fibers by blending synthetic polymer solutions with selected extracellular matrix proteins. The correct choice of polymer also makes the usage of toxic and denaturing solvents unnecessary. In this study we have investigated the production process for and the suitability of protein/PEO fibers, specifically collagen I/PEO, as cell culture substrates.