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|Title:||Recharacterization of the RGC-5 retinal ganglion cell line|
|Author:||Van Bergen, N.|
|Citation:||Investigative Ophthalmology & Visual Science, 2009; 50(9):4267-4272|
|Publisher:||Assoc Research Vision Ophthalmology Inc|
|Nicole J. Van Bergen, John P. M. Wood, Glyn Chidlow, Ian A. Trounce, Robert J. Casson, Won-Kyu Ju, Robert N. Weinreb, and Jonathan G. Crowston|
|Abstract:||PURPOSE: The transformed RGC-5 retinal ganglion cell line is used widely in glaucoma research. Increased resistance to glutamate was noted in published literature and led to the recharacterization of the RGC-5 cell line. METHODS: Characterization of the RGC-5 cell line was performed by sequencing of a region of the nuclear Thy1 gene and mitochondrial DNA sequencing of a region of the d-loop and tRNA(Phe) gene. Marker expression was examined in undifferentiated cells, and cells differentiated with 50 microg/mL succinyl concanavalin A (S Con A) for 3 days. Glutamate sensitivity was examined in undifferentiated and S Con A differentiated cells by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay after 24-hours of glutamate treatment. RESULTS: RGC-5 cells were found to be of mouse (Mus musculus), not rat (Rattus norvegicus), origin by mitochondrial and nuclear DNA analyses. RGC-5 DNA sequenced in a second laboratory was subsequently found to be of M. musculus origin. Cells stained positively for the neuronal markers beta-tubulin and PGP9.5 and for the microtubule-associated protein tau, but not for known markers of ganglion cells such as neurofilaments or Thy1.2, suggesting that they likely represented a lineage of mouse neuronal precursor cells. Differentiation with S Con A did not increase RGC-5 sensitivity to glutamate excitotoxicity or increase the expression of retinal or ganglion cell marker proteins. CONCLUSIONS: Investigators using cells designated as RGC-5 should confirm the species to be of rat origin and retinal-specific marker expression before considering their use as retinal ganglion-like cells.|
|Keywords:||Retinal Ganglion Cells; Cell Line, Transformed; Animals; Mice; Rats; Glutamic Acid; Concanavalin A; Biological Markers; Fluorescent Antibody Technique, Indirect; Cell Differentiation|
|Appears in Collections:||Opthalmology & Visual Sciences publications|
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