Fibrous scaffold and physical stimuli are known to enhance muscle development. In this study, we fabricated polycaprolactone (PCL) fibers in aligned or random arrangements by electrospinning. PCL fibers were spun towards either spaced collector or flat plate collector. C2C12 muscle cells (mouse myoblasts) were seeded on scaffolds coated with fibronectin (10μg/ml) and let grow until cells became confluent. Cells were then cultured for a week under horse serum condition. Immunofluorescence for skeletal myosin heavy chain, an indicator for muscle differentiation, showed enhanced differentiation of muscle cells on aligned fibers when compared to ones on random condition.. Synergetic effects of fiber alignments and DC electric field were further investigated to propose an appropriate condition for muscle differentiation for tissue engineering.