Isolation and Culture of Primary Retinal Müller Cells from Sprague-Dawley (SD) Rats.

Retinal Müller cells (RMCs) play a crucial role in providing structural support and regulating various functions within the retina. As a core component of the retinal microenvironment, RMCs perform several vital functions. Through their abundant ion channels, ligands, receptors, transmembrane transporters, and enzyme systems, these cells contribute to neurotransmitter and trophic factor secretion, regulate retinal metabolism, and maintain water-ion homeostasis. Notably, RMCs have recently been identified as a significant source of endogenous retinal regenerative stem cells, offering novel therapeutic targets for the treatment of retinal degenerative diseases. Therefore, studying RMCs is essential for understanding the pathological mechanisms underlying retinal disorders. This study systematically establishes a standardized experimental protocol that includes trypsin digestion and purification of primary RMCs, morphological observation using inverted optical microscopy and hematoxylin and eosin (HE) staining, specific protein identification through immunofluorescence staining, and cell purity analysis via flow cytometry. This protocol serves as a valuable reference for both basic research and clinical applications related to RMCs, supporting the exploration of their mechanisms in retinal diseases and advancing the development of therapeutic strategies.