April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Generation Of Photoreceptor Phenotypes From Induced Nrl-gfp Mouse Hair Follicle-derived Epithelial Stem Cells
Author Affiliations & Notes
  • Catherine Jomary
    Cell Lab Ltd, Southampton, United Kingdom
  • Stephen E. Jones
    Biochemistry, King’s College London, London, United Kingdom
  • Mandeep Singh
    Nuffield Laboratory of Ophthalmology, University of Oxford, Oxford, United Kingdom
  • Paul Wong
    Ophthalmology, Emory University, Atlanta, Georgia
  • Robert MacLaren
    Nuffield Laboratory of Ophthalmology, University of Oxford, Oxford, United Kingdom
  • Footnotes
    Commercial Relationships  Catherine Jomary, Cell Lab Ltd (E); Stephen E. Jones, None; Mandeep Singh, None; Paul Wong, None; Robert MacLaren, None
  • Footnotes
    Support  Health Foundation
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 3181. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Catherine Jomary, Stephen E. Jones, Mandeep Singh, Paul Wong, Robert MacLaren; Generation Of Photoreceptor Phenotypes From Induced Nrl-gfp Mouse Hair Follicle-derived Epithelial Stem Cells. Invest. Ophthalmol. Vis. Sci. 2011;52(14):3181.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: : Hair follicles provide a readily accessible potential donor source of autologous cells for photoreceptor transplantation, if these cells could be reprogrammed. Previous work with the nrl.gfp transgenic mouse has shown that the post-mitotic rod precursor represents the optimal stage for transplantation into a degenerate retina The aim of the present study was to induce rod photoreceptor precursors from hair follicle-derived epithelial stem cells harvested from the nrl.gfp mouse. This was achieved by modulating the microenvironment in vitro using retina-specific conditioned media and a retinal explant co-culture system.

Methods: : Epithelial stem cells were isolated from the hair follicle bulge region of whiskers from 2 week old nrl.gfp mice by mechanical dissection, enriched by clonal expansion, and subcultured in retina-conditioned media or with retinal explants. Changes in the mRNA and protein 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 GMP phosphodiesterase) were evaluated by immunocytochemistry, Western blotting, and reverse transcription-polymerase chain reaction.

Results: : Isolated stem cells from the nrl.gfp mouse hair follicle bulges were successfully expanded by clonal growth. Media derived from cultured retinal cells or retinal explant co-culture systems were found to be effective in inducing differentiation of skin stem cells into rod photoreceptor-like cells. Expression of stem cell markers of proliferation and pluripotency was decreased. Significantly, cells expressed bright GFP de novo together with photoreceptor-specific markers.

Conclusions: : The present study demonstrates that hair follicle epithelial stem cells can be reprogrammed into photoreceptor-like cells when exposed to a retina-specific microenvironment. Since the induced expression of GFP is driven by the nrl transcription factor, these cells may be at the optimal developmental stage for retinal repair by autologous transplantation.

Keywords: photoreceptors • retina • gene/expression 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×