The Secretome of Alginate-Encapsulated Limbal Epithelial Stem Cells Modulates Corneal Epithelial Cell Proliferation
نویسندگان
چکیده
Limbal epithelial stem cells may ameliorate limbal stem cell deficiency through secretion of therapeutic proteins, delivered to the cornea in a controlled manner using hydrogels. In the present study the secretome of alginate-encapsulated limbal epithelial stem cells is investigated. Conditioned medium was generated from limbal epithelial stem cells encapsulated in 1.2% (w/v) calcium alginate gels. Conditioned medium proteins separated by 1-D gel electrophoresis were visualized by silver staining. Proteins of interest including secreted protein acidic and rich in cysteine, profilin-1, and galectin-1 were identified by immunoblotting. The effect of conditioned medium (from alginate-encapsulated limbal epithelial stem cells) on corneal epithelial cell proliferation was quantified and shown to significantly inhibit (P≤0.05) their growth. As secreted protein acidic and rich in cysteine was previously reported to attenuate proliferation of epithelial cells, this protein may be responsible, at least in part, for inhibition of corneal epithelial cell proliferation. We conclude that limbal epithelial stem cells encapsulated in alginate gels may regulate corneal epithelialisation through secretion of inhibitory proteins.
منابع مشابه
Variations of the Normal Human Limbal Stem Cells Detected by In Vivo Confocal Microscopy
Background To report normal variations of the limbal structures using in vivo laser scanning confocal microscopy. Methods: This was a retrospective study of fourteen eyes from 11 healthy individuals. Confocal imaging of cornea and limbus was performed using Heidelberg Retina Tomograph III Rostock Corneal Module. Results: The typical structure of the palisades of Vogt (POV) was detected ...
متن کاملComparison of Ultra Structure and Gene Expression of Cultured Limbal Stem Cells and Fresh Conjunctival, Limbal and Corneal Tissues
Purpose: The present study intends to show the characteristics of cultured limbal stem cell (CLSCs) and to compare them with normal Conjunctival (C), Limbal (L) and Cornea (K) tissues. Materials and Methods: The expressions of a set of genes potentially involved in differentiation and stemness function of limbal stem cells were assessed in freshly prepared limbal, corneal, and conjunctival tis...
متن کاملOxidized alginate hydrogels as niche environments for corneal epithelial cells
Chemical and biochemical modification of hydrogels is one strategy to create physiological constructs that maintain cell function. The aim of this study was to apply oxidised alginate hydrogels as a basis for development of a biomimetic niche for limbal epithelial stem cells that may be applied to treating corneal dysfunction. The stem phenotype of bovine limbal epithelial cells (LEC) and the v...
متن کاملCorneal epithelial stem cells at the limbus: looking at some old problems from a new angle.
Corneal epithelium is traditionally thought to be a self-sufficient, self-renewing tissue implying that its stem cells are located in its basal cell layer. Recent studies indicate however that corneal epithelial stem cells reside in the basal layer of peripheral cornea in the limbal zone, and that corneal and conjunctival epithelia represent distinct cell lineages. These ideas are supported by ...
متن کاملKeratin 13 is a more specific marker of conjunctival epithelium than keratin 19
Introduction To evaluate the expression patterns of cytokeratin (K) 12, 13, and 19 in normal epithelium of the human ocular surface to determine whether K13 could be used as a marker for conjunctival epithelium. Methods: Total RNA was isolated from the human conjunctiva and central cornea. Those transcripts that had threefolds or higher expression levels in the conjunctiva than the cornea wer...
متن کامل