Abstract Purpose The goal of current research was to develop the material for scaffold preparation providing optimal conditions for the corneal epithelium cell culturing and to determine their physicochemical and biological properties. The scaffolds, which could be applied in ophthalmology should fulfill some requirements such as biocompatibility, biodegradability, restorability, and also should have adequate mechanical strength Methods A membrane was obtained from chitosan‐polysaccharide derived from chitin. Chitosan is nontoxic,biologically inert,stable in the natural environmental. It has found numerous applications in biomedical field,e.g.3Dscaffolds,fibers,membranes, hydrogels. This materials have shown support for adhesion,proliferation,expression of cells. The chitosan supports will be crosslinked with genipin‐product from the Gardenia jasminoides. The genipin is more than 5000 times less cytotoxic than typical chemical cross‐linking reagents (glutaraldehyde). The mechanical properties of the supports obtained will be controlled mainly by the addition other biopolymers Results The in vitro cell culture experiments with corneal epithelium cells have indicated that a membrane prepared from chitosan blends provided the regular stratified growth of the epithelium cells, good surface covering and increased number of the cell layers. Some of chitosan membrane is comparable with that of the amniotic membrane which is currently recommended for clinical applications Conclusion Novel polymeric hydrogel scaffolds for corneal epithelium cell culturing based on blends of chitosan with some other biopolymers such as hydroxypropylcellulose, collagen and elastin crosslinked with genipin, a natural substance, were prepared