title: Effect of Müller cell co-culture on in vitro permeability of bovine retinal vascular endothelium in normoxic and hypoxic conditions.
abstract: Müller cell dysfunction may contribute to the early pathological changes associated with conditions such as diabetes, that cause breakdown of the blood-retinal barrier. In this study we used an in vitro model of the blood-retinal barrier to investigate Müller cell effects on retinal vascular endothelial cell monolayer permeability under normoxic (20% oxygen) and hypoxic (1% oxygen) conditions. Second passage bovine retinal capillary endothelial cells were co-cultured with retinal Müller cells on opposite sides of a 0.4 microm pore size polycarbonate Transwell filter or in medium that was continually conditioned by Müller cells. Permeability changes were observed for up to 24h of hypoxia by measurement of [(3)H]-inulin and [(14)C]-albumin flux across the endothelial cell monolayer. Endothelial cell barrier function was enhanced by co-culturing with Müller cells under normoxic conditions. Under hypoxic conditions however, the barrier was significantly impaired after 12h of co-culture with Müller cells. These results shed more light on the trophic effect of Müller cells on the blood-retinal barrier, suggesting a critical role in the maintenance and regulation of the barrier in health and during disease.
authors.affiliation: Save Sight Institute, Department of Clinical Ophthalmology, University of Sydney, 8 Macqarie Street, Sydney, NSW 2001, Australia. marina@eye.usyd.edu.au
authors.forename: Marina
authors.initials: M
authors.lastname: Tretiach
authors.forename: Michele C
authors.initials: MC
authors.lastname: Madigan
authors.forename: Li
authors.initials: L
authors.lastname: Wen
authors.forename: Mark C
authors.initials: MC
authors.lastname: Gillies
mesh_terms: D000418:Albumins / Q000378:metabolism / Q000493:pharmacokinetics; D000818:Animals; D001813:Blood-Retinal Barrier / Q000502:physiology*; D002417:Cattle; D002450:Cell Communication / Q000502:physiology; D002463:Cell Membrane Permeability / Q000502:physiology*; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology*; D000860:Hypoxia / Q000503:physiopathology*; D007444:Inulin / Q000378:metabolism / Q000493:pharmacokinetics; D008954:Models, Biological; D009457:Neuroglia / Q000166:cytology / Q000502:physiology*; D012160:Retina / Q000166:cytology / Q000502:physiology*
chemical_list: D000418:Albumins; D007444:Inulin