Generation of a Biomimetic Substitute of the Corneal Limbus Using Decellularized Scaffolds Sánchez Porras, David Caro Magdaleno, Manuel González Gallardo, Carmen García García, Óscar Darío Garzón Bello, Ingrid Johanna Carriel Araya, Víctor Campos Sánchez, Fernando Alaminos Mingorance, Miguel Corneal limbus Decellularized xenograft Recellularization Mesenchymal stem cells Patients with severe limbal damage and limbal stem cell deficiency are a therapeutic challenge. We evaluated four decellularization protocols applied to the full-thickness and half-thickness porcine limbus, and we used two cell types to recellularize the decellularized limbi. The results demonstrated that all protocols achieved efficient decellularization. However, the method that best preserved the transparency and composition of the limbus extracellular matrix was the use of 0.1% SDS applied to the half-thickness limbus. Recellularization with the limbal epithelial cell line SIRC and human adipose-derived mesenchymal stem cells (hADSCs) was able to generate a stratified epithelium able to express the limbal markers p63, pancytokeratin, and crystallin Z from day 7 in the case of SIRC and after 14–21 days of induction when hADSCs were used. Laminin and collagen IV expression was detected at the basal lamina of both cell types at days 14 and 21 of follow-up. Compared with control native limbi, tissues recellularized with SIRC showed adequate picrosirius red and alcian blue staining intensity, whereas limbi containing hADSCs showed normal collagen staining intensity. These preliminary results suggested that the limbal substitutes generated in this work share important similarities with the native limbus and could be potentially useful in the future. 2021-11-08T09:03:23Z 2021-11-08T09:03:23Z 2021 info:eu-repo/semantics/article Sánchez-Porras, D.; Caro-Magdaleno, M.; González-Gallardo, C.; García-García, Ó .D.; Garzón, I.; Carriel, V.; Campos, F.; Alaminos, M. Generation of a Biomimetic Substitute of the Corneal Limbus Using Decellularized Scaffolds. Pharmaceutics 2021, 13, 1718. https://doi.org/10.3390/ pharmaceutics13101718 http://hdl.handle.net/10481/71349 10.3390/pharmaceutics13101718 eng http://creativecommons.org/licenses/by/3.0/es/ info:eu-repo/semantics/openAccess Atribución 3.0 España MDPI