Quantifying Retinal Pigment Epithelial Phagocytosis and Lipofuscin Accumulation

April 14, 2023

Improved Lipofuscin Models and Quantification of Outer Segment Phagocytosis Capacity in Highly Polarized Human Retinal Pigment Epithelial Cultures (Video) | JoVE

02:52 min

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Preparation of Photo-Oxidized Outer Segment Tips and Fragments (Video) | JoVE

03:57 min

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Characterization of the Lipofuscin-Like Granule Spectrum and Composition (Video) | JoVE

02:05 min

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Assessing the Effects of Lipofuscin-Like Granules on RPE Phagocytosis: Total Consumptive Capacity (Video) | JoVE

02:17 min

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Qitao Zhang, Gillian Autterson, Jason M. L. Miller²

¹Kellogg Eye Center,University of Michigan, Ann Arbor, ²Cellular and Molecular Biology Program,University of Michigan, Ann Arbor

Summary

This protocol describes a lipofuscin accumulation model in highly differentiated and polarized human retinal pigment epithelial (RPE) cultures and an improved outer segment (OS) phagocytosis assay to detect the total OS consumption/degradation capacity of the RPE. These methods overcome the limitations of previous lipofuscin models and classical pulse-chase outer segment phagocytosis assays.