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C. Jomary, S. E. Jones, A. J. Lotery; Differentiation of Rat Hair Follicle Stem Cells Into Photoreceptor Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):1232.
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The aim of this study was to evaluate the potential of rat hair follicle stem cells to transdifferentiate into photoreceptor-like cells, following modulation of the microenvironment using retinal-specific conditions or after genetic modification using the Crx transcription factor.
Epithelial stem cells were isolated from the hair follicle bulge region by mechanical dissection, enriched by clonal expansion, and either subcultured in retinal-conditioned media, or genetically modified by electroporation to express exogenous epitope-tagged murine Crx. Changes in the expression of stem cell markers (homeodomain transcription factor Pax6, POU transcription factor Oct3/4) and putative skin stem cell markers (K15, alpha 6 integrin), neuronal markers (nestin, neuron-specific class III ß-tubulin and neurofilament), and photoreceptor-specific markers (rhodopsin, cyclic nucleotide-gated cation channel-3, blue-cone opsin, cyclic (c)GMP phosphodiesterase) were evaluated by immunocytochemistry, Western blotting, and quantitative reverse transcription-polymerase chain reaction.
Isolated stem cells from the hair follicle bulge were successfully expanded by clonal growth without a feeder layer. Both media derived from cultured retinal cells and exogenous expression of Crx by genetic modification were found to be effective in inducing a photoreceptor -like phenotype. Expression of stem cell markers of proliferation and pluripotency was decreased. Concomitantly, expression of neuronal and photoreceptor-specific markers was up-regulated.
The present study suggests that rat hair follicle epithelial stem cells are capable of differentiation into photoreceptor phenotype cells ex vivo when either exposed to retinal-specific microenvironment or genetically modified with Crx. The present study extends our previous findings that exogenous Crx expression can induce mouse and human retina-derived stem cells into functional photoreceptor cells and is consistent with the notion that Crx has a broadly-applicable ability to promote differentiation of such cells into photoreceptor phenotypes.
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