May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Transformed Rat Retinal Ganglion Cells (RGC-5) Develop a Differentiated Morphology Upon Co-Culture With Human Non-Pigmented Ciliary Epithelial (HNPE) Cells
Author Affiliations & Notes
  • R.M. Dauphin
    Pharmacology and Neuroscience, UNT Health Science Ctr, Fort Worth, TX, United States
  • R.R. Krishnamoorthy
    Pharmacology and Neuroscience, UNT Health Science Ctr, Fort Worth, TX, United States
  • G. Prasanna
    Pharmacology and Neuroscience, UNT Health Science Ctr, Fort Worth, TX, United States
  • T. Yorio
    Pharmacology and Neuroscience, UNT Health Science Ctr, Fort Worth, TX, United States
  • Footnotes
    Commercial Relationships  R.M. Dauphin, None; R.R. Krishnamoorthy, None; G. Prasanna, None; T. Yorio, None.
  • Footnotes
    Support  EY11979
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 2249. doi:
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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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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : 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.

Keywords: ganglion cells • retinal culture • ciliary body 
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