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Type: Journal article
Title: A comparison of differentiation protocols for RGC-5 cells
Author: Wood, J.
Chidlow, G.
Tran, T.
Crowston, J.
Casson, R.
Citation: Investigative Ophthalmology & Visual Science, 2010; 51(7):3774-3783
Publisher: Assoc Research Vision Ophthalmology Inc
Issue Date: 2010
ISSN: 0146-0404
Statement of
John P. M. Wood, Glyn Chidlow, Tuyet Tran, Jonathan G. Crowston, and Robert J. Casson
Abstract: Purpose: Although the RGC-5 cell line is widely used in retinal ganglion cell (RGC) research, recent data have raised questions about the nature of these cells. The authors performed a systematic analysis of RGC-5 cells to determine which RGC or neuronal markers are expressed after treatment with known differentiating agents, thus providing further insight into the nature of these cells and assisting in defining their future use. Methods: RGC-5 cells were treated for 5 days with staurosporine (STSN; 316 nM), trichostatin A (TSA; 500 nM), or succinylconcanavalin A (sConA; 50 _g/mL), after which they were assayed for specific marker antigen/mRNA expression. Treated cells were also assayed for excitotoxic responsiveness. Results: Neither treated nor untreated RGC-5 cells expressed any specific RGC marker mRNAs or proteins (Brn-3, neurofilaments, Thy-1) or calbindin, calretinin, synaptophysin, PKC_, or glial fibrillary acidic protein. However, control RGC-5 cells did express the neuronal markers tau, _III-tubulin, microtubule-associated protein (MAP)-1b, MAP2, and PGP9.5. Although treatment with sConA had no effect on the expression of these markers, STSN and (dose dependently) TSA increased their expression and induced excitotoxic responsiveness. All cells, treated or not, expressed high levels of nestin but no other progenitor cell markers. All cells also expressed cone-specific, but not rod-specific, opsin indicative of cone photoreceptor lineage. Conclusions: RGC-5 cells expressed neuronal, but not RGCspecific, markers that were dose dependently upregulated by TSA. Hence, TSA provided the best tested means to terminally differentiate the cells to a neuronal phenotype from a precursor- like lineage.
Keywords: Retinal Ganglion Cells; Cell Line; Animals; Mice; Hydroxamic Acids; Staurosporine; Intermediate Filament Proteins; Concanavalin A; Nerve Tissue Proteins; RNA, Messenger; Biological Markers; Electrophoresis, Polyacrylamide Gel; Reverse Transcriptase Polymerase Chain Reaction; Cell Differentiation; Cell Lineage; Dose-Response Relationship, Drug; Opsins
Rights: Copyright © Association for Research in Vision and Ophthalmology
RMID: 0020100258
DOI: 10.1167/iovs.09-4305
Grant ID:
Appears in Collections:Medicine publications

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