title: Analysis of photoreceptor outer segment phagocytosis by RPE cells in culture.
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 describe three different and complementary methods to quantify total POS uptake by RPE cells and to discriminate surface-bound from engulfed POS.
authors.affiliation: Department of Biological Sciences, Fordham University, Bronx, NY, USA.
authors.forename: Yingyu
authors.initials: Y
authors.lastname: Mao
authors.forename: Silvia C
authors.initials: SC
authors.lastname: Finnemann
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D015151:Immunoblotting / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D055355:Opsins / Q000032:analysis; D010587:Phagocytosis; D055214:Retinal Photoreceptor Cell Outer Segment / Q000648:ultrastructure*; D055213:Retinal Pigment Epithelium / Q000166:cytology*; D013552:Swine
chemical_list: D055355:Opsins