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dc.contributor.authorVan Bergen, N.en
dc.contributor.authorWood, J.en
dc.contributor.authorChidlow, G.en
dc.contributor.authorTrounce, I.en
dc.contributor.authorCasson, R.en
dc.contributor.authorJu, W.en
dc.contributor.authorWeinreb, R.en
dc.contributor.authorCrowston, J.en
dc.identifier.citationInvestigative Ophthalmology & Visual Science, 2009; 50(9):4267-4272en
dc.description.abstractPURPOSE: 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.en
dc.description.statementofresponsibilityNicole J. Van Bergen, John P. M. Wood, Glyn Chidlow, Ian A. Trounce, Robert J. Casson, Won-Kyu Ju, Robert N. Weinreb, and Jonathan G. Crowstonen
dc.publisherAssoc Research Vision Ophthalmology Incen
dc.subjectRetinal Ganglion Cells; Cell Line, Transformed; Animals; Mice; Rats; Glutamic Acid; Concanavalin A; Biological Markers; Fluorescent Antibody Technique, Indirect; Cell Differentiationen
dc.titleRecharacterization of the RGC-5 retinal ganglion cell lineen
dc.typeJournal articleen
pubs.library.collectionOpthalmology & Visual Sciences publicationsen
dc.identifier.orcidChidlow, G. [0000-0001-7371-0239]en
dc.identifier.orcidCasson, R. [0000-0003-2822-4076]en
Appears in Collections:Opthalmology & Visual Sciences publications

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