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J. Amaral, P. Becerra; Diffusion of Pigment Epithelium Derived Factor (PEDF) Through the Outer Blood Retinal Barrier . Invest. Ophthalmol. Vis. Sci. 2003;44(13):391.
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Purpose: PEDF is a potent natural antiangiogenic protein which has been shown to inhibit retinal neovascularization when administered systemically. The purpose of the present study is to evaluate the diffusion of PEDF through monkey retinal pigment epithelial (mRPE) cells monolayer in culture. Methods: Early passage Rhesus monkey RPE cells were grown to confluence in 12mm polycarbonate membrane inserts (0.4um pore size) loaded in 12 well plates. Confluence was followed by measuring the transepithelial resistance. Fluorescinated horse radish peroxidase (Fl-HRP, 500ng/ml) was used as negative control. Fl-PEDF was prepared by conjugating human recombinant PEDF with Fluorescein and fluorescinated protein (10nM) were added to each basal and apical compartments of membrane inserts.Fluorescence was measured using a plate reader. Results: Culture RPE cells in membrane inserts had a transepithelial resistance ranging between 120-140 ohms. In inserts with Fl-PEDF on both compartments the apical:basal ratio increased to 1:1.25 by 22 hs.In inserts with Fl-PEDF only in the basal compartment the apical:basal ratio was 1:1.23. Conclusions: These results suggest that PEDF can pass through mRPE cells in vitro and that this could be the mechanism buy which PEDF crossing the outer blood retinal barrier exerts its antiangiogenic effects following systemic administration.
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