Extreme ultraviolet (EUV) surface modification of polytetrafluoroethylene (PTFE) was performed in order to enhance the degree of biocompatibility. Polymer samples were irradiated by different number of EUV shots using a laser–plasma based EUV source in the presence of nitrogen gas. The physical and chemical properties of EUV modified PTFE samples were studied using Atomic Force Microscopy, X-ray photoelectron spectroscopy and water contact angle (WCA) methods. Pronounced wall type micro and nano-structures appeared on the EUV treated polymer surfaces resulting in increased surface roughness and hydrophobicity. Stronger cell adhesion and good cell morphology were observed on EUV modified surfaces by in-vitro cell culture studies performed using L929 fibroblasts.