PT  - JOURNAL ARTICLE
AU  - Fangyuan Gao
AU  - Emily Tom
AU  - Dorota Skowronska-Krawczyk
TI  - A novel quantification method for RPE phagocytosis using a VLC-PUFA-based strategy
AID  - 10.1101/2022.07.02.498584
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.07.02.498584
4099  - http://biorxiv.org/content/early/2022/07/03/2022.07.02.498584.short
4100  - http://biorxiv.org/content/early/2022/07/03/2022.07.02.498584.full
AB  - The retinal pigment epithelium (RPE) lies adjacent to the photoreceptors and is responsible for the engulfment and degradation of shed photoreceptor outer segment fragments (POS) through receptor-mediated phagocytosis. Phagocytosis of POS is critical for maintaining photoreceptor function and is a key indicator of RPE functionality. The current methods to study RPE phagocytosis are not compatible with direct quantification with high sensitivity that can be applied in a high-throughput manner. To overcome some of these limitations, we have developed a novel VLC-PUFA-based approach for evaluating RPE phagocytic activity in primary bovine RPE and ARPE-19 cells and validated its results by traditional methods. This new approach can be used to detect the dynamic process of phagocytosis at varying POS concentrations and incubation times and offers a robust, quantifiable, and unbiased assay for high-throughput screening.Competing Interest StatementDS-K is a scientific advisor of Visgenx, Inc.