Abstract
Retinal pigment epithelial (RPE) cells are among the most actively phagocytic cells in nature. Primary RPE and stable RPE cell lines provide experimental model systems that possess the same phagocytic machinery as RPE in situ. Upon experimental challenge with isolated photoreceptor outer segment fragments (POS), these cells promptly and efficiently recognize, bind, internalize, and digest POS. Here, we describe experimental procedures to isolate POS from porcine eyes and to feed POS to RPE cells in culture. Furthermore, we provide experimental protocols to synchronize the POS binding and engulfment steps of phagocytosis. Finally, we describe three different and complementary methods to quantify total POS uptake by RPE cells and to discriminate surface-bound from engulfed POS.
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This work was supported by NIH grant EY026215.
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Mazzoni, F., Mao, Y., Finnemann, S.C. (2019). Advanced Analysis of Photoreceptor Outer Segment Phagocytosis by RPE Cells in Culture. In: Weber, B.H.F., Langmann, T. (eds) Retinal Degeneration. Methods in Molecular Biology, vol 1834. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8669-9_7
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DOI: https://doi.org/10.1007/978-1-4939-8669-9_7
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-8668-2
Online ISBN: 978-1-4939-8669-9
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