Objective: To culture astrocytes of optic nerve and to establish the cell lines for further study of healing process after optic nerve trauma.Methods: Optic nerve astrocytes of infantile and adults with sudden death were cultured by tissue inoculation or tissue digestion with 0. 25% Trypsin and 0. 06% EDTA. The second and fourth passage cells were stained with HE and and- GFAP, S-100 protein, Vi-mentin, and CD34 antibodies.Results: The trypsinized astrocytes of infantile optic nerve reached confluence in 7 days, but the astrocytes of adults weren’t successfully cultured. The cultured cells were in polygonal shape with processes; the cytoplasm was abundant and pink; the cells had light-blue nuclei. These cells were positive in GFAP, S-100 protein and vimentin staining, and negative in CD34 staining.Conclusions: The results showed that astrocytes of infantile optic nerve can be successfully cultured and trypsinization is a better method than tissue inoculation. The culture of infantile astrocytes is easie Objective: To culture astrocytes of optic nerve and to establish the cell lines for further study of healing process after optic nerve trauma. Methods: Optic nerve astrocytes of infantile and adults with sudden death were cultured by tissue inoculation or tissue digestion with 0.25% Trypsin and 0. 06% EDTA. The second and fourth passage cells were stained with HE and and- GFAP, S-100 protein, Vi-mentin, and CD34 antibodies. Results: The trypsinized astrocytes of infantile optic nerve reach confluence in 7 days , but the astrocytes of adults were not successfully cultured. The cultured cells were in polygonal shape with processes; the cytoplasm was abundant and pink; the cells had light-blue nuclei. These cells were positive in GFAP, S-100 protein and vimentin staining, and negative in CD34 staining. Conclusions: The results showed that astrocytes of infantile optic nerve can be successfully cultured and trypsinization is a better method than tissue inoculation. The culture of infantile a strocytes is easie