Low-glucose enhances keratocyte-characteristic phenotype from corneal stromal cells in serum-free conditions
نویسندگان
چکیده
The avascular cornea is a uniquely-isolated organ, with its stroma constituting a nutrient-poor environment. Consequently, the availability of metabolites such as glucose to corneal stromal cells is considerably reduced compared with other tissues, or indeed with media commonly used to culture these cells in vitro. However, the role of glucose in the behaviour of human corneal keratocytes has been overlooked. As such, we sought to investigate the effects of low-glucose formulations on the phenotype of human corneal stromal cells. Cells cultured in low-glucose were able to survive for extended periods when compared to high-glucose, serum-free conditions. Furthermore, low-glucose enhanced their reversal to a keratocyte-characteristic phenotype. Specifically, cells within low-glucose medium assumed dendritic morphologies, with bean-shaped condensed nuclei, absence of alpha-smooth muscle actin or stress fibres, and a corresponding reduction in migratory and contractile activities when compared with high-glucose, serum-free conditions. Moreover, cells within low-glucose uniquely recovered the ability to express a robust keratocyte-characteristic marker, CD34, while still expressing elevated levels of other representative phenotypic markers such as keratocan, lumican, ALDH1A1, and ALDH3A1. These results indicate that low-glucose enhances keratocyte-characteristic phenotype above and beyond established media formulations and thus has important implications for corneal biology in health and disease.
منابع مشابه
Application of retinoic acid improves form and function of tissue engineered corneal construct
Retinoic acid has recently been shown to control the phenotype and extracellular matrix composition of corneal stromal cells cultured in vitro as monolayers. This study set out to investigate the effects of retinoic acid on human corneal keratocytes within a 3D environment. Human corneal keratocytes were encapsulated in collagen gels, which were subsequently compressed under load, and cultured ...
متن کاملRetinoic Acid Improves Keratocyte Phenotype in a 3D Model of the Corneal Stroma
Program Number: 1300 Presentation Time: 8:30 AM–8:45 AM Retinoic Acid Improves Keratocyte Phenotype in a 3D Model of the Corneal Stroma Ricardo M. Gouveia, Fadhilah Z. Abidin, Che J. Connon. Institute of Genetic Medicine, Newcastle University, Newcastle-upon-Tyne, United Kingdom. Purpose: Retinoic Acid (RA) is a metabolite of vitamin A fundamental for the correct function and development of the...
متن کاملAdipose-derived stem cells differentiate to keratocytes in vitro
PURPOSE Adipose-derived stem cells (ADSC) are an abundant population of adult stem cells with the potential to differentiate into several specialized tissue types, including neural and neural crest-derived cells. This study sought to determine if ADSC express keratocyte-specific phenotypic markers when cultured under conditions inducing differentiation of corneal stromal stem cells to keratocyt...
متن کاملSerum-free spheroid culture of mouse corneal keratocytes.
PURPOSE To develop a serum-free mass culture system for mouse keratocytes. METHODS Corneas of C57BL6/J mice were enzyme digested after the epithelium and endothelium were removed. Stromal cells were cultured in serum-free DMEM/F12 (1:1) containing epidermal growth factor (EGF), fibroblast growth factor 2 (FGF2), and B27 supplement. Primary spheres were dissociated by trypsin and subcultured a...
متن کاملSecretion and organization of a cornea-like tissue in vitro by stem cells from human corneal stroma.
PURPOSE To investigate the potential of human corneal stromal stem cells to assume a keratocyte phenotype and to organize extracellular matrix (ECM) in vitro similar to corneal stromal tissue. METHODS Human corneal stromal stem cells (hCSSC) were isolated as side population cells by flow cytometry. Cloned hCSSC were cultured as free-floating pellets in serum-free media for 3 weeks. Gene expre...
متن کامل