Cell Biology, Neurobiology, and Anatomy, Medical College of Wisconsin, Milwaukee, WI, USA; Ophthalmology and Visual Sciences, Medical College of Wisconsin, Milwaukee, WI, USA.
Department of Ophthalmology and Visual Sciences, Heersink School of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA.
STAR Protoc. 2022 Dec 16;3(4):101758. doi: 10.1016/j.xpro.2022.101758. Epub 2022 Oct 12.
Primary culture and long-term maintenance of primary retinal pigment epithelium (RPE) is a useful model system for the study of ocular pathologies such as age-related macular degeneration. Here, we detail the steps for the isolation and long-term culture of primary porcine RPE. We also describe steps for cryoprotecting, cryosectioning, and interrogating with immunofluorescence and histochemistry RPE cells grown on transwell membranes. These techniques can be used in histological studies to detect sub-RPE deposits. For complete details on the use and execution of this protocol, please refer to Pilgrim et al., (2017).
原代培养和长期维持原代视网膜色素上皮（RPE）是研究眼部疾病（如年龄相关性黄斑变性）的有用模型系统。在这里，我们详细介绍了分离和长期培养原代猪 RPE 的步骤。我们还描述了在 Transwell 膜上生长的 RPE 细胞进行冷冻保护、冷冻切片和免疫荧光及组织化学检测的步骤。这些技术可用于组织学研究，以检测 RPE 下沉积物。有关此方案的使用和执行的完整详细信息，请参阅 Pilgrim 等人（2017 年）。
