Abstract
Purpose :
Previous studies have demonstrated tissue-related changes in various scleral proteins with myopia, primarily in animal models. However, the cellular characteristics of healthy human scleral fibroblasts are not fully established. In this study, human scleral fibroblasts were obtained and grown in cell culture to establish the baseline characteristics of healthy cells, including proliferation and protein and collagen content.
Methods :
Commercially available human scleral fibroblasts were cultured in Lifeline FibroLife medium with 2% serum. Cells were maintained at 37 degrees Celsius and 5% CO2. To verify cellular identity, the cells were characterized by immunocytochemistry of vimentin and actin filaments. Cells were also assessed for the presence of glycosaminoglycans, proteoglycans, and transforming growth factor-β2 by immunocytochemistry. An EdU assay was performed to evaluate cell proliferation.
Results :
Immunocytochemistry against fibroblast markers showed characteristic staining patterns of vimentin and actin filaments. Collagen I, lumican, versican and hyaluronic acid were immunopositive, whereas decorin, chondroitin sulphate, keratan sulphate, biglycan, and transforming growth factor-β2 were immunonegative. Cells grown from passages 3 to 7 were morphologically stable and 57.50 ± 5.33% were EdU positive.
Conclusions :
This study verified the cellular identity of commercially available human scleral fibroblasts and assessed their proliferation characteristics. These baseline data provide information regarding relevant proteoglycans, glycosaminoglycans, and structural proteins that may be involved in myopic scleral remodelling and will aid our future studies seeking to create an in vitro environment that mimics the microenvironment of myopic scleral fibroblasts.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.