title: Chitosan-gelatin biopolymers as carrier substrata for limbal epithelial stem cells.
abstract: The aim of this work was to evaluate semi-synthetic biopolymers based on chitosan (CH) and gelatin (G) as potential in vitro carrier substrata for human limbal epithelial cells (hLECs). To that end, human corneal epithelial cells (HCE) were cultured onto different CH-G membranes. None of the polymers were cytotoxic and cell proliferation was higher when CH was functionalized with G. Expression levels of corneal epithelial markers (K3, K12, E-caherin, desmoplakin, and zonula occludens (ZO)-1) were better maintained in HCE cells grown on CH-G 20:80 membranes than other proportions. Consequently, CH-G 20:80 was chosen for the subsequent expansion of hLECs. Cells derived from limbal explants were successfully expanded on CH-G 20:80 membranes using a culture medium lacking components of non-human animal origin. The expression levels found for corneal (K3 and K12) and limbal epithelial stem cells (K15) specific markers were similar to or higher than those found in limbal cells grown onto the control substratum. Our results demonstrate that CH-G 20:80 membranes are suitable for the expansion and maintenance of stem cells derived from the limbal niche. These results strongly support the use of polymers as alternative substrata for the transplantation of cultivated limbal cells onto the ocular surface.
authors.affiliation: IOBA (Institute of Applied Ophthalmobiology), University of Valladolid, Valladolid, Spain, adelamatas@ioba.med.uva.es.
authors.forename: Ana
authors.initials: A
authors.lastname: de la Mata
authors.forename: Teresa
authors.initials: T
authors.lastname: Nieto-Miguel
authors.forename: Marina
authors.initials: M
authors.lastname: López-Paniagua
authors.forename: Sara
authors.initials: S
authors.lastname: Galindo
authors.forename: María Rosa
authors.initials: MR
authors.lastname: Aguilar
authors.forename: Luis
authors.initials: L
authors.lastname: García-Fernández
authors.forename: Sandra
authors.initials: S
authors.lastname: Gonzalo
authors.forename: Blanca
authors.initials: B
authors.lastname: Vázquez
authors.forename: Julio San
authors.initials: JS
authors.lastname: Román
authors.forename: Rosa María
authors.initials: RM
authors.lastname: Corrales
authors.forename: Margarita
authors.initials: M
authors.lastname: Calonge
mesh_terms: D001704:Biopolymers / Q000737:chemistry*; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002470:Cell Survival; D048271:Chitosan / Q000737:chemistry*; D003315:Cornea / Q000473:pathology; D003470:Culture Media / Q000737:chemistry; D019573:Epithelium, Corneal / Q000166:cytology*; D005780:Gelatin / Q000737:chemistry*; D006801:Humans; D008422:Materials Testing; D011108:Polymers / Q000737:chemistry; D013234:Stem Cells / Q000166:cytology*; D054457:Tissue Scaffolds
chemical_list: D001704:Biopolymers; D003470:Culture Media; D011108:Polymers; D005780:Gelatin; D048271:Chitosan
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