Protocol for the isolation, culturing, and evaluation of primary Müller glial cells from pig retina as cellular models in health and disease

Müller glial cells maintain retinal homeostasis and provide structural and metabolic support. The pig eye closely resembles the human eye in morphology and physiology, offering strong translational potential. Here, we present a protocol for isolating and culturing primary Müller glial cells from pig retina to study their function in health and disease. We describe steps for eyeball dissection, retinal isolation, photoreceptor cell depletion, and retinal digestion. We then detail procedures for the selection, culture, and characterization of Müller glial cells. • Steps for Müller glial cell isolation from porcine retina • Instructions for the selection of Müller glial cells via D-sorbitol media • Procedures for freezing and long-time storage of primary porcine Müller glial cells • Guidance on fluorescence microscopy analysis for primary porcine Müller glial cultures Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Müller glial cells maintain retinal homeostasis and provide structural and metabolic support. The pig eye closely resembles the human eye in morphology and physiology, offering strong translational potential. Here, we present a protocol for isolating and culturing primary Müller glial cells from pig retina to study their function in health and disease. We describe steps for eyeball dissection, retinal isolation, photoreceptor cell depletion, and retinal digestion. We then detail procedures for the selection, culture, and characterization of Müller glial cells.