Abstract
Purpose :
Innovation in the application of alternative stem-cell based therapies, specifically, with respect to tissue engineering offers opportunities to improve treatment of corneal disorders. In this study, we obtain baseline data to test the hypothesis that non-sulphated and/or minimally sulphated chondroitin sulphate proteoglycans (CSPGs) are able to regulate the proliferation and differentiation of embryonic chick corneal stromal keratocytes, perhaps promoting a stem cell phenotype, as is believed to be the case in cartilage.
Methods :
Primary chick corneal keratocytes were isolated at embryonic day 18 (E18) and expanded in culture. Immunohistochemical analysis of matrix secretion (pro-collagen type I, collagen type I, collagen type III, keratocan, and keratan sulphate) and the expression of putative stromal stem cell markers (connexion 43 (CX43), PAX6, BMI-1, ABCG2 and K3) were assessed. In situ expression of these stromal markers was evaluated at developmental stages between E10 and E20.
Results :
In cell cultures, a strong immunohistochemical signal was observed for collagen I, pro-collagen I, keratocan and keratan sulphate and lesser signal for collagen III. In corneal tissue sections, no stromal expression of PAX6 was observed while strong, diffuse expression of CX43 was noted throughout the developmental stages.
Conclusions :
The present findings establish a baseline from which future treatment of keratocytes with varying patterns of sulphated CSPGs can be investigated, with the overarching aim to establish a desirable in vitro environment that promotes a stem cell phenotype.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.