Controlled extracellular chemical and topographical cues can generate physicochemical changes that influence the proliferation and differentiation of neural cells; external electrical stimulation (ES) via conductive bioelectrodes can promote neural differentiation by increasing neurite outgrowth. Rat pheochromocytoma (PC12) cells are used as a neuron‐like model cells to explore the possibility of using a well‐designed poly(ethylene oxide) (PEO)/poly(3,4‐ethylenedioxythiophene):polystyrenesulfonate (PEDOT:PSS) blend solution to fabricate functional bioelectrodes presenting biological fouling/antifouling surfaces, thereby regulating the cell adhesion, proliferation, and differentiation properties. In this study, it is found that a flat PEO/PEDOT:PSS composite film fabricates through spin‐coating, operates through a contact repulsion mechanism that limits cell attachment and proliferation; in contrast, the aligned and random PEO/PEDOT:PSS nanofibers fabricates through electrospinning promoted neuron adhesion efficiently and allows manipulation of the cell morphology. Furthermore, ES of PC12 cells is performed to investigate the influence of the conductive random and aligned PEO/PEDOT:PSS composite nanofiber mats on the enhancement of neurite outgrowth, as well as the relative gene expression of Nestin, Tuj1, and MAP2. Therefore, combining this unique PEDOT:PSS blend solution with various fabrication processes appears to be a facile approach toward bioelectronic interface coatings displaying tunable surface properties for manipulating the cellular behavior of neurons during ES.