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I. A. Trounce, N. J. Van Bergen, J. P. Wood, G. Chidlow, R. J. Casson, J. G. Crowston; Re-Characterisation of the RGC-5 Retinal Ganglion Cell Line. Invest. Ophthalmol. Vis. Sci. 2009;50(13):2786.
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© ARVO (1962-2015); The Authors (2016-present)
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 re-characterisation of the RGC-5 cell line.
Characterisation of the RGC-5 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 tRNAPhe gene. Marker expression was examined in undifferentiated cells, and cells differentiated with 50ug/ml Succinyl Concanavalin A (S Con A) for 3 days. Glutamate sensitivity was examined in undifferentiated and S Con A differentiated cells by MTT assay after 24 hours exposure to glutamate
RGC-5 cells were found to be of mouse (Mus musculus), not rat (Rattus norvegicus) origin by both mitochondrial and nuclear DNA analysis. RGC-5 DNA sourced from two further laboratories was also found to be of Mus musculus origin. Cells stained positively for the neuronal markers β-tubulin and PGP9.5, as well as for the microtubule-associated protein, tau, but not for known markers of ganglion cells such as neurofilaments or Thy1.2 suggesting that these cells likely represent a linage of mouse neuronal precursor cells. Differentiation with S Con A did not increase RGC-5 sensitivity to glutamate excitotoxicity, or increase expression of retinal or ganglion cell marker proteins.
We suggest that 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.
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