Abstract

PurposeRGC-5 cells undergo differentiation into a neuronal phenotype with low concentrations of staurosporine. Although the RGC-5 cell line was initially thought to be of retinal ganglion cell origin, recent evidence suggests that the RGC-5 line could have been the result of contamination with 661W mouse cone photoreceptor cells. This raised the possibility that a cone photoreceptor cell line could be multipotent and could be differentiated to a neuronal phenotype.Methods661W and RGC-5 cells, non-neuronal retinal astrocytes, retinal endothelial cells, retinal pericytes, M21 melanoma cells, K562 chronic myelogenous leukemia cells, and Daudi Burkitt lymphoma cells, were differentiated with staurosporine. The resulting morphology was quantitated using NeuronJ with respect to neurite counts and topology.ResultsTreatment with staurosporine induced similar-appearing morphological differentiation in both 661W and RGC-5 cells. The following measures were not significantly different between 661W and RGC-5 cells: number of neurites per cell, total neurite field length, number of neurite branch points, and cell viability. Neuronal-like differentiation was not observed in the other cell lines tested.Conclusions661W and RGC-5 cells have virtually identical and distinctive morphology when differentiated with low concentrations of staurosporine. This result demonstrates that a retinal neuronal precursor cell with cone photoreceptor lineage can be differentiated to express a neuronal morphology.

Highlights

  • The RGC-5 cell line was produced as a continuously proliferating model of rat retinal ganglion cells (RGCs)

  • 661W and RGC-5 cells have virtually identical and distinctive morphology when differentiated with low concentrations of staurosporine

  • This result demonstrates that a retinal neuronal precursor cell with cone photoreceptor lineage can be differentiated to express a neuronal morphology

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Summary

Methods

Staurosporine (isolated from Streptomyces staurosporeus) was obtained from Alexis Biochemical (San Diego, CA). The number of neurites extended after staurosporine treatment did not significantly differ between 661W and RGC-5 cells at any concentration tested (Fig 1). This data was used to assess total neurite field length and number of neurite branch points, a measure of neurite field complexity, for each condition For both of these measures, the differentiation response of 661W and RGC-5 cells did not significantly differ. For both cell types, maximum neurite field length and maximum neurite branch points were observed after treatment with 316 nM staurosporine. To test non-retinal continuous cell lines, the response of the M21 melanoma, K562 chronic myelogenous leukemia, and Daudi Burkitt lymphoma cells to staurosporine was tested None of these cell lines exhibited morphological differentiation after treatment with 316 nM staurosporine (Fig 3)

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