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Abstract Summary Cornea is the transparent part that represents ⅙ of the outer layer of the eyeball. Opacification of cornea could be resulted from infection, trauma, autoimmune disease, and hereditary causes. Loss of corneal clearance is irreversible in most of cases. Despite of high demands for corneal tissue transplantation in patients with corneal disorders, there is a significant shortage in corneal tissue donation. For this reason, continuous researches and studies discussed the role of tissue engineering and regenerative medicine to find an alternative for tissue donation. Collagen gel is one of new biomaterials that could be used as a substrate for corneal tissue regeneration. Based on characteristic and mechanical properties of human cornea, it was found that limbal epithelial stem cells tend to keep their phenotype in the basal layer of corneal limbus. Limbus is found to be softer in its mechanical properties than central part of cornea. Aim of the work: The aim of this research was to study the hypothesis that the normal function of an adult tissue stem cell niche is controlled by the mechanical properties of the extracellular matrix that surrounds it. This was achieved via expansion of human primary corneal epithelial stem cells using substrates of differing stiffness. Materials and methods: Donated corneas: Four cadaverous human corneas were used for studying histology and immunohistochemistry of human corneas. Eight donated cadaverous human |