3D cell culture formats more closely resemble tissue architecture complexitythan 2D systems, which are lacking most of the cell–cell andcell–microenvironment interactions of the in vivo milieu. Scaffold-basedsystems integrating natural biomaterials are extensively employed in tissueengineering to improve cell survival and outgrowth, by providing the chemicaland physical cues of the natural extracellular matrix (ECM). Using thefreeze–drying technique, porous 3D composite scaffolds consisting ofpoly(3,4-ethylene-dioxythiophene) doped with polystyrene sulfonate(PEDOT:PSS), containing ECM components (i.e., collagen, hyaluronic acid,and laminin) are engineered for hosting neuronal cells. The resulting scaffoldsexhibit a highly porous microstructure and good conductivity, determined byscanning electron microscopy and electrochemical impedance spectroscopy,respectively. These supports boast excellent mechanical stability and wateruptake capacity, making them ideal candidates for cell infiltration. SH-SY5Yhuman neuroblastoma cells show enhanced cell survival and proliferation inthe presence of ECM compared to PEDOT:PSS alone. Whole-cell patch-clamprecordings acquired from differentiated SHSY5Y cells in the scaffoldsdemonstrate that ECM constituents promote neuronal differentiation in situ.These findings reinforce the usability of 3D conducting supports asengineered highly biomimetic and functional in vitro tissue-like platforms fordrug or disease modeling.