Quantification of pigment epithelium-derived factor (PEDF) in an ex vivo coculture of retinal pigment epithelium cells and neuroretina

Salvatore Di Lauro1,2, Maria Teresa Garcia-Gutierrez1, Ivan Fernandez-Bueno1,3,4

1. Instituto Universitario de Oftalmobiología Aplicada (IOBA), Universidad de Valladolid, Valladolid, Spain
2. Departamento de Oftalmología, Hospital Clinico Universitario de Valladolid, Valladolid, Spain
3. Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y Leon, Valladolid, Spain
4. Red Temática de Investigación Cooperativa en Salud (RETICS), Oftared, Instituto de Salud Carlos III, Valladolid, Spain

Abstract

Purpose: To quantify the pigment epithelium-derived factor (PEDF) levels in a coculture model of physically separated neuroretina and retinal pigment epithelium (RPE) cells, and to point out its potential role in neuroretinal maintenance. Methods: RPE cells and neuroretina explants were isolated from porcine eyes. RPE cells were expanded and seeded on the bottom of Transwell® culture inserts. Neuroretina explants were cultured alone (controls) on Transwell® culture membranes or supplemented with RPE cells in the same wells but physically separated by the culture membrane, during 9 days. PEDF concentration in the culture medium at 3, 5, 7 and 9 days of culture was determined by enzyme-linked immunosorbent assays (ELISA) specific for porcine samples. Mean statistical analysis were performed with Pair-wise Student’s-tests. Results: Culture medium collected from neuroretina cultures without and with RPE cells contained detectable levels of PEDF in both conditions and at all evaluated time-points. At 3, 5, and 7 days and through the whole time of culture PEDF concentration was significantly higher in cocultures with RPE cells (p<0.05). Conclusions: RPE cells neuroprotective role may be linked to the beneficial effects of neurotrophic factors, such as PEDF, secreted or induced by RPE cells during co-culture.
Keywords: Ex vivo model; Retinal pigment epithelium (RPE); Neuroretina; Pigment Epithelium-Derived Factor (PEDF); Neuroprotection
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Publication
Ms. Ref. No.: A0201008     Author-Reviewer    
Received: 12/09/2019  
Accepted: 06/11/2019   
Published: 15/04/2020 

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