Gum tragacanth is one of the most widely used natural gums in food, medicine, cosmetics and personal care products, and its use as polysaccharide‐based scaffolds in tissue engineering applications has attracted great attention in recent years. The fabrication of pure gum tragacanth as a scaffold poses many challenges because of the high viscosity, poor mechanical properties and repulsive interaction between the polyanions. To overcome these, facilitate the formation of scaffolds and improve their final properties, chitosan and gum tragacanth were used together as natural, biocompatible and biodegradable polysaccharides. The scaffolds based on chitosan and gum tragacanth were successfully fabricated through cryotropic gelation and were characterized using different chemical, morphological, mechanical and biocompatibility analyses. All cryogel scaffolds exhibited a porous structure with an average diameter of 96.56–30.21 μm, exhibiting high liquid absorption capacity, appropriate mechanical stability and controlled degradation behavior. According to the biocompatibility results, mouse embryonic fibroblast cells adhered well to the scaffolds and achieved high viability. The results are also discussed in the light of their potential usefulness as a scaffold for tissue engineering applications. © 2022 Society of Industrial Chemistry.