The purpose of this research was to investigate the effect of glucose concentration on thermally induced cross-linking in electrospun gelatin fibres. Scaffolds were prepared from both type A and type B gelatin and used in cell culture experiments. Fourier transform infrared spectroscopy (FTIR) was used to monitor the cross-linking process. Fibre morphology and structure of obtained scaffolds were analysed by scanning electron microscopy (SEM). Scaffolds with different glucose concentrations were tested for possible leaching of glucose and enzyme mediated degradation. In order to evaluate the ability to support cell growth and proliferation, fibroblasts were seeded on scaffolds with different glucose content and analysed after culturing for one day (24 h) and seven days.