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https://hdl.handle.net/2440/51311
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Type: | Journal article |
Title: | Recharacterization of the RGC-5 retinal ganglion cell line |
Author: | Van Bergen, N. Wood, J. Chidlow, G. Trounce, I. Casson, R. Ju, W. Weinreb, R. Crowston, J. |
Citation: | Investigative Ophthalmology and Visual Science, 2009; 50(9):4267-4272 |
Publisher: | Assoc Research Vision Ophthalmology Inc |
Issue Date: | 2009 |
ISSN: | 0146-0404 1552-5783 |
Statement of Responsibility: | 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: | <h4>Purpose</h4>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.<h4>Methods</h4>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.<h4>Results</h4>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.<h4>Conclusions</h4>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 Fluorescent Antibody Technique, Indirect Cell Differentiation Biomarkers |
DOI: | 10.1167/iovs.09-3484 |
Published version: | http://dx.doi.org/10.1167/iovs.09-3484 |
Appears in Collections: | Aurora harvest Opthalmology & Visual Sciences publications |
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