Seeding a layer of cells at specific depths within scaffolds is an important optimization parameter for bi-layer skin models. The work presented investigated the effect of fiber diameter and its mechanical property on the depth of cell seeding for electro-spun fiber scaffold. Polycaprolactone (PCL) is used to generate scaffolds that are submicron (400nm) to micron (1100nm) using electro-spinning. 3T3 fibroblasts were seeded on the electro-spun fiber scaffold mat of 50-70 microns thickness in this study. In order to investigate the effect of fiber diameter on cell migration, first, the electrospun fiber scaffold was studied for variation of mechanical properties as a function of fiber diameters. Atomic force microscopy (AFM) was used to investigate the Young’s modulus (E) values as a function of fiber diameter. It was identified that as the fiber diameter increases, the Young’s modulus values decreases considerably from 1.1GPa to 200MPa. The variation in E is correlated with cell seeding depth as a function of vacuum pressure. A higher E value led to a lower depth of cell seeding (closer to the surface) indicating that nanofibrous scaffolds offer larger resistance to cell movement compared to microfibrous scaffolds.