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Kailin Tian, Ben Davis, Li Guo, Nidhi Aggarwal, Daren Hanumunthadu, Lisa Turner, Shereen Nizari, M Francesca Cordeiro; A Comparison of the RGC-5 Cell Line and Primary Murine Retinal Cultures Under Oxidative Stress. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):4957.
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© ARVO (1962-2015); The Authors (2016-present)
The RGC-5 cell line has been used in a wide variety of studies to investigate retinal ganglion cell (RGC) activity since it was first developed and characterized in 2001. However, recent data have raised serious questions regarding the true nature of these cells. Although now highly likely the RGC-5 line is not of gangion cell origin, the question remains as to how similar these cells behave to primary retinal cultures. Continued characterisation of the RGC-5 line is important as this line has been used in over 200 studies and at present, no other RGC line exists. RGC-5 is reported to express neuronal markers (Thy1 and Brn3a). The present study sought to compare cellular response to oxidative stress between RGC-5 and primary mixed retinal cultures. Oxidative stress is known to contribute to pathology of a variety of retinal diseases, including optic neuropathy and glaucoma. The aim of this study was to compare the behaviour of RGC-5 with primary retinal culture under oxidative stress, induced by Dimethylsulfoxide (DMSO),a known cell insult which inhibits the antioxidant function, and paraquat, a known electron acceptor in redox and radical reactions.
Cell viability experiments were performed in the presence of different concentrations of DMSO and paraquat in RGC-5 cell line and murine mixed retinal cultures. Cell survival was measured after 24h exposure to cytotoxic insults using the Alamar Blue viability assay. IC50 values were compared in each case.
Similar toxicity was observed between RGC-5 and primary mixed retinal cultures induced by DMSO and paraquat. The IC50 for RGC-5 and primary murine mixed retinal cultures were 2.474% and 2.027% respectively for DMSO, 136.5μM and 330μM respectively for paraquat.
Similar toxicity profiles were observed between RGC-5 and mixed murine retinal cultures in response to oxidative stress in vitro. Increased resistance to Paraquat insult may be due to the presence of glial cells in mixed retinal cultures. These are well documented to buffer retinal oxidative stress. This study suggests, although far from ideal, the response of the RGC-5 cell line to oxidative stress is comparable to mixed retinal cultures and can be used as in-vitro models for retina disorders.
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