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RC Nayak, MG K Kwok, BM Stramer, PF Olson, ME Fini; The 3G5 Cell Surface Ganglioside is a Marker of Corneal Keratocytes in vivo and in vitro . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1701.
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Purpose: As there is a paucity of markers of keratocytes and as keratocytes and capillary pericytes both have mesenchymal origins, we evaluated the anti-pericyte monoclonal antibody, 3G5, for use as a keratocyte marker. Methods: 3G5 antigen expression was studied by immunofluorescence on frozen sections of cornea and on tissue cultured stromal cells. Results: The 3G5 antigen is a ganglioside, a sialic acid containing glycosphingolipid. This antigen migrates between the ganglioside markers G m1 and G m2 on thin layer chromatography but is otherwise uncharacterized. We found that 3G5 stained frozen sections of human, rat and rabbit cornea but not mouse cornea. The staining pattern followed the distribution of keratocytes in the stroma but did not stain the epithelium or endothelium. The stain was found on long thin processes and was often punctate. 3G5 staining was assessed in rabbit cornea with full thickness punch wounds at day 20 of healing. In these full thickness wounds, the fibroblastic mass in the center of the wound was 3G5 negative, the margins of the wound showed increased 3G5 staining, areas distant from the wound showed a normal 3G5 staining pattern. In human keratocyte cultures that were sub-confluent 3G5 stained 0 % of the cells. Almost 100% of the human cells that were maintained at confluence for two weeks were 3G5 positive. Tissue cultured human neonatal foreskin fibroblasts and adult human dermal fibroblasts do not express the 3G5 antigen. 100% of tissue cultured rabbit keratocytes were 3G5 positive regardless of whether they were isolated and cultivated in serum free or serum containing medium. Conclusion: The 3G5 antigen is an excellent marker of corneal keratocytes in vivo and in vitro. These results also indicate that tissue cultured keratocytes can be distinguished from fibroblasts by 3G5 antigen expression.
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