ABSTRACT Chitosan‐based scaffolds are widely studied in tissue regeneration because of their biocompatibility and biodegradability. Scaffolds are obtained by different techniques and can be modified with other polymers allowing controlling their properties. This article discusses the assembling of three‐dimensional chitosan porous scaffolds blended with gelatin. Gelatin was used to enhance cells attachment due to the presence of cell adhesion motifs, while improving mechanical strength. 2,5‐dimethoxy‐2,5‐dihydrofurane (DHF) was used as the crosslinking agent, because it allowed to control the reaction kinetics through temperature, time and DHF concentration. The results indicate that scaffolds morphology, pore sizes and distribution, compressive moduli and biodegradation in vitro with lysozyme, can be customized with variations of gelatin content and crosslinking degree. Scaffolds were neither cytotoxic nor genotoxic for human keratinocytes, exhibiting cell–substrate interactions. Our findings demonstrated that chitosan–gelatin scaffolds crosslinked with DHF, as a new crosslinking agent, are suitable in tissue engineering applications. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133 , 43814.