Abstract

Previous studies have suggested a critical interdependence of choroidal endothelial cells (EC) and retinal pigment epithelial (RPE) cells, which are separated by the Bruch's membrane (BrM), for the maintenance of retinal function. We developed a long‐term coculture system to evaluate the effect of EC‐RPE interactions on cell behavior and matrix deposition. Human RPE cells and primary human EC were seeded on opposite sides of polyester transwells and cultured for up to 4 weeks in low serum medium. Cells and extracellular matrix deposition were analyzed using viability assay, transepithelial resistance, H&E staining, immunohistochemistry, and qPCR. Coculture with RPE sustained EC survival and proliferation in low serum for up to 2 weeks. Presence of EC induced RPE expression of matrix‐associated and visual cycle proteins. Interestingly, expression of PEDF and Thrombospondin‐1 by RPE were also significantly upregulated in coculture condition, suggesting that RPE acquired an anti‐angiogenic status. TEM and immunohistochemistry analysis revealed the accumulation of BrM‐like material on the basal side of cocultured RPE. RPE differentiation and induced deposition of certain matrix‐associated proteins in coculture further emphasizes the significance of RPE‐EC interactions in retinal homeostasis. This work was supported by the NIH Director's New Innovator Award 1DP2OD006649

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call

Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.