The nanofibrous structure containing protein and polysaccharide has good potential in tissue engineering. The present work aims to study the role of chitosan in gelatin/chitosan nanofibrous scaffolds fabricated through electrospinning process under optimized condition. The performance of chitosan in gelatin/chitosan nanofibrous scaffolds was evaluated by mechanical tests, scanning electron microscopy (SEM), Fourier transform infrared (FTIR) and in vitro cell culture on scaffolds with different gelatin/chitosan blend ratios. To assay the influence of chitosan ratio on biocompatibility of the electrospun gelatin/chitosan scaffolds for skin tissue engineering, the culturing of the human dermal fibroblast cells (HDF) on nanofibers in terms of attachment, morphology and proliferation was evaluated. Morphological observation showed that HDF cells were attached and spread well on highly porous gelatin/chitosan nanofibrous scaffolds displaying spindle-like shapes and stretching. The fibrous morphologies of electrospun gelatin/chitosan scaffolds in culture medium were maintained during 7 days. Cell proliferation on electrospun gelatin/chitosan scaffolds was quantified by MTS assay, which revealed the positive effect of chitosan content (around 30%) as well as the nanofibrous structure on the biocompatibility (cell proliferation and attachment) of substrates.