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MS Good, R Varma; Anti-apoptotic Genes (bcl-2, bag-1, p35) Increase the Survival of Retinal Ganglion Cells in Culture . Invest. Ophthalmol. Vis. Sci. 2002;43(13):2188.
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Purpose: Retinal ganglion cells (RGCs) damaged in glaucoma die through apoptosis. Introducing anti-apoptotic genes into RGCs may prevent this death. This study was performed to determine whether the simultaneous introduction and supplementation of cell survival genes (bcl-2, bag-1, p35) into cultured RGCs using adenoviral vectors would delay apoptotic RGC death. Methods: RGCs were isolated from post-natal day 8 Sprague Dawley rats using the two step immunopanning technique. Purified cells were plated at a density of 8,000 cells per well of a 96 well plate and maintained in serum free medium with growth factors. RGCs were then infected with an adenoviral vector driven by a CMV promoter containing the bcl-2, bag-1, and p35 genes. A second group of RGCs was supplemented with the cell survival genes each day. Control cells were incubated with an adenoviral vector driven by a CMV promoter containing the lacZ gene. RGC survival was assessed as the percentage of viable cells on a specific day compared to the number of cells plated at day 0. Results: RGC survival increased significantly in cells transfected with all three cell survival genes. The greatest effect on RGC survival was seen after simultaneous transfection and supplementation with all three genes.
% RGC Survival View OriginalDownload SlideView OriginalDownload Slide Conclusion: Simultaneous transfection and supplementation with all three cell survival genes significantly increases survival of RGCs in culture.
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