Mao Yingyu, Finnemann Silvia C
Department of Biological Sciences, Fordham University, Bronx, NY, USA.
Methods Mol Biol. 2013;935:285-95. doi: 10.1007/978-1-62703-080-9_20.
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.
视网膜色素上皮（RPE）细胞是自然界中最活跃的吞噬细胞之一。原代RPE细胞和稳定的RPE细胞系提供了与原位RPE具有相同吞噬机制的实验模型系统。在用分离的光感受器外段片段（POS）进行实验性刺激时，这些细胞能迅速有效地识别、结合、内化并消化POS。在此，我们描述了从猪眼中分离POS并将其投喂给培养中的RPE细胞的实验步骤。此外，我们还描述了三种不同且互补的方法，用于量化RPE细胞对总POS的摄取，并区分表面结合的POS和被吞噬的POS。
