Generation of a biomimetic human artificial cornea model using Wharton's Jelly mesenchymal stem cells

Purpose: Human Wharton's jelly stem cells (HWJSCs) derived from the umbilical cord have unique advantages due to their stemness differentiation capability at in vitro and in vivo levels. Recently, it has been demonstrated that HWJSCs are able to differentiate into epithelial-like cells of skin and oral mucosa and they could be considered as exceptional candidates for tissue regeneration. In this work, we have demonstrated for the first time the capability of HWJSCs to differentiate in vitro into cornea epithelial-like cells using a three dimensional model based on fibrin-agarose human bioengineered corneas. Methods: First, primary cell cultures of HWJSCs, corneal epithelial cells and corneal keratocytes were cultured and three-dimensional ortotypic and heterotypic human cornea models were generated. Results: Our results demonstrated that HWJSCs were able to differentiate to corneal epithelial-like cells by tissue engineering strategies with similar results to native cornea. Apart from this, heterotypical corneas using HWJSCs revealed adequate differentiation levels of the epithelium and stroma, being similar to ortotypic corneas and native corneas for the expression of epithelial markers (CK3/12, PKG, ZO1 and CX43) and extracellular matrix components (proteoglycans, collagen, elastic and reticular fibers). Moreover, immunofluorecence analysis confirmed the presence of crystallins, Cry-αA, Cry-αB, Cry-β, Cry-ζ with low expression of Cry-βγ3 and Cry-λ1 key proteins involved in cornea transparency, in both models. Conclusions: All these findings suggest that HWJSCs could be considered as an alternative cell source for cornea regeneration and may offer a solution to patients with limbus stem cell deficiency