Conductive core–sheath nanofibers are prepared by a combination of electrospinning and aqueous polymerization. Specifically, nanofibers electrospun from poly(ε-caprolactone) (PCL) and poly(L-lactide) (PLA) are employed as templates to generate uniform sheaths of polypyrrole (PPy) by in-situ polymerization. These conductive core–sheath nanofibers offer a unique system to study the synergistic effect of different cues on neurite outgrowth in vitro. It is found that explanted dorsal root ganglia (DRG) adhere well to the conductive core–sheath nanofibers and generate neurites across the surface when there is a nerve growth factor in the medium. Furthermore, the neurites can be oriented along one direction and enhanced by 82% in terms of maximum length when uniaxially aligned conductive core–sheath nanofibers are compared with their random counterparts. Electrical stimulation, when applied through the mats of conductive core–sheath nanofibers, is found to further increase the maximum length of neurites for random and aligned samples by 83% and 47%, respectively, relative to the controls without electrical stimulation. Together these results suggest the potential use of the conductive core–sheath nanofibers as scaffolds in applications such as neural tissue engineering.