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R.M. Dauphin, R.R. Krishnamoorthy, G. Prasanna, T. Yorio; Transformed Rat Retinal Ganglion Cells (RGC-5) Develop a Differentiated Morphology Upon Co-Culture With Human Non-Pigmented Ciliary Epithelial (HNPE) Cells . Invest. Ophthalmol. Vis. Sci. 2003;44(13):2249.
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Purpose: : The purpose of this study was to differentiate a virally transformed rat retinal ganglion cell line (RGC-5). This cell line exhibits several marker proteins characteristic of retinal ganglion cells, however it has a neuroepithelial morphology. Inducing morphological changes consistent with a neuronal phenotype may be one indication of differentiation of the RGC-5 cells. Methods: HNPE cells were seeded on collagen inserts in DMEM complete medium and grown to confluence for 3 days. RGC-5 cells were seeded in 6 well plates and after 1 hour the inserts containing the HNPE cells were added on top of the RGC-5 layer. After 5 days of co-culture the wells were observed by light microscopy to determine if any morphological changes had occurred. Currently we are treating RGC-5 with various doses of PEDF and other trophic factors as potential differentiating agents. Results: RGC-5 cells developed a more neuron-like appearance after incubation with HNPE cells for 5 days. The RGC-5 cells exhibited several neurites and a long thin axon extending from the cell bodies. The control RGC-5 cells, without co-culture, had a more flattened epithelial-like appearance, with short fat extensions. Conclusions: Differentiation factors secreted from HNPE cells induce a morphological change in the RGC-5 cells. One well characterized factor secreted by HNPE cells is pigmented epithelial-derived factor (PEDF). PEDF has neurotrophic, neuroprotective, and neurite growth-promoting activity and could be one of the factors contributing to the differentiation of RGC-5 cells. A differentiated population of RGC-5 cells could be useful to study various aspects of retinal ganglion cell biology.
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