Purified mouse retinal ganglion cell culture

Objective: To establish a culture system for purified mouse retinal ganglion cells (RGC) culture in order to lay a foundation for the in vitro study of RGC. Methods: It was a experiment study. Eight to twelve C57BL/6 mice on postnatal day 4 to 6 were used. The retinas were dissected and dissociated enzymatically to make a suspension of single cells. The retinal suspension was incubated in rat anti-mouse-macrophage antiserum for 5 minutes and incubated on a 100 mm anti-rat IgG panning plate at room temperature for 30 min twice. The nonadherent cells were removed with the suspension and placed on the Thy-1.2 panning plate at room temperature. After 45 min, plates were washed 6-10 times with phosphate-buffered saline and swirled moderately to dislodge nonadherent cells. Trypsin (0.125% solution) dissociation was used to remove adherent cells from the plate. Then cells were spun and resuspended in neurobasal medium containing some neurotrophic factors. The cell suspension was implanted in 24 well culture plates and cultured under 37°C in an incubator with 5% C0 ₂. Results: Most of the cells adhered to the plate after 24 hours in culture and showed dendrites at different lengths. The dendrites grew longer with time. Most of the RGC can survive more than two weeks. Conclusion: Monoclonal antibody to the Thy-1.2 antigen can be used to purify the mice RGC.