A Novel Primary Porcine Retinal Pigment Epithelium Cell Model with Preserved Properties

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Purpose To establish an ethical, reliable and expandable retinal pigment epithelial (RPE) cell model with maintained RPE properties compatible with multifarious assays.Methods RPE cells from abattoir-obtained porcine eyes were cultured under various conditions. Morphology, RPE cell-specific protein markers (RPE-65, CRALBP) and the tight junction marker ZO-1 were analysed by phase-contrast microscopy, immunocytochemistry and western blot, and transepithelial electrical resistance (TEER) was determined to assess barrier function.Results The porcine RPE cells (pRPE) were best established using TrypLE Express, 10% fetal bovine serum (FBS) supplemented high-glucose media and subculturing at semi-confluency. The pRPE cells maintained epithelioid morphology with ZO-1 positive tight junctions at the cell-to-cell borders, ability to establish proper barrier function (TEERmax: 346/375 Ω⋅cm2 at passage I/passage VI) and expressed CRALBP and RPE-65 for several passages. The RPE characteristics decreased and disappeared with transdifferentiation.Conclusions This work describes, for the first time, a pRPE cell model that exhibits preserved RPE properties for several passages on cell culture plastic plates. Though RPE characteristics were maintained for at least 6 passages, the reduced CRALBP and RPE-65 with passaging emphasize that lower passage cells are advantageous to utilise, and that morphology, barrier function and ZO-1 localisation cannot be solely employed as a quality measure of RPE identity. Pigs are phylogenetically similar to humans, including similar physiology, anatomy and immune system. Therefore, porcine RPE cells constitute a relevant model system for studying human eye diseases, such as AMD.

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