Ocular Cell Monolayers Cultured on Biodegradable Substrates

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Abstract
The aim of this study was to culture retinal pigment epithelial (RPE) and corneal endothelial cells on biodegradable substrates for future use in monolayer transplantation in the eye. The biodegradable polymers, poly-L-lactic (PLLA) and poly-DL-lactic-co-glycolic acid (85:15) (PLGA) (both of molecular weight 105 kd) were the biomaterials used. All materials were seeded with either pig/human retinal pigment epithelial cells or rabbit corneal endothelial cells and were maintained in tissue culture conditions. Upon confluency, the cell density was calculated and cell viability determined. All monolayers were stained with phalloidin-rhodamine for F-actin and antibodies to the tight junction (zonula occludens) protein, ZOi, to demonstrate the presence of tight junctions. The final cell density of human RPE monolayers on PLLA films was 2950 cells/mm2 (±185). The final cell density of pig RPE on PLLA and PLGA film was 2350 cells/mm2 (±152 and 178, respectively). Rabbit corneal endothelial cells had a final cell density of 2650 cells/mm2 (±164). F-actin staining revealed a circumferential ring of actin filaments in all of the cells grown on substrates. ZOi immunohistochemistry demonstrated staining along the lateral cell borders of all cell types. The successful culture of retinal pigment epithelial and corneal endothelial monolayers on these substrates may have potential for transplanting cell monolayers in the eye to improve vision.