Primary culture of rabbit retinal Mūeller cells

Objective Mailer cells play critical roles in maintaining retinal physiologic function and are involve in some of pathogenesis in many retinal diseases. Therefore, the cultivation of retinal Mūeller cells is value to establish a platform for the studies of Mūeller cells in responses to the environment changes in physiologic and pathologic condition. Aim of present study was to establish an effective method to isolate and culture rabbit retinal Mūeller cells. Methods Mūeller cells were dissociated from young rabbit retinas using an enzymatic digestion technique and primary cultured in 10% FBS DMEM/F12. Shaking and washing process were used to allow the neurons detaching from Mūeller cells. The cell cultures in 3rd passage were confirmed as Mūeller cell by microscope observation, immunocytochemical staining with glial fibrillary acidic protein （GFAP） antibody and transmission electron microscope analysis. Results Scattering attached cells were seen in culture dishes by 24 hours after primary culture. 72 hours latter,the numbers of attached cells were gradually increased. The cultured cells grew and reached confluence in 20 -25 days. The cultured Mūeller cells showed a flatten tripolar shape with 95% of cells showing positive immunohistochemical reaction for glial fibrillary acidic protein. Mitochondria, glycogen particles and intermediate filaments （8 - 10nm） were seen in a monolayer of the cultured cells using transmission electron microscopy,indicating the characters of retinal Mūeller cells the cultured cells. Conclusion Rabbit retinal Mueller cells can be isolated using an enzymatic digestion technique successfully and purified by shaking and washing. Key words: cell culture;  retinal glial cells; Mūeller cells