Vitro culture and immunohistochemical identification of astrocytes of infantile optic nerve.
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 anti-GFAP, S-100 protein, Vimentin, 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 easier than that of adult astrocytes. Immunohistochemistry were used to determine the source and type of those cultured cells.
Zheng
J
J
Zhongshan Ophthalmic Center, Sun Yat-sen University of Medical Sciences and National Ophthalmological Laboratories, Ministry of Public Health, China.
Lan
Y
Y
Zhang
J
J
Guo
Y
Y
Luo
Y
Y
D000328:Adult; D001253:Astrocytes / Q000166:cytology*; D002478:Cells, Cultured; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D007223:Infant; D009900:Optic Nerve / Q000166:cytology*; D009418:S100 Proteins / Q000032:analysis; D014746:Vimentin / Q000032:analysis*
D005904:Glial Fibrillary Acidic Protein; D009418:S100 Proteins; D014746:Vimentin