May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Sox2 Is Required for Maintaining the Stem Cell Characteristics of Human Müller Stem Cells From the Adult Retina
Author Affiliations & Notes
  • B. Bhatia
    Pathology and Cell Biology, UCL Institute of Ophthalmology, London, United Kingdom
  • S. Singhal
    Pathology and Cell Biology, UCL Institute of Ophthalmology, London, United Kingdom
    Moorfields Eye Hospital, London, United Kingdom
  • J. S. Ellis
    Pathology and Cell Biology, UCL Institute of Ophthalmology, London, United Kingdom
  • P. T. Khaw
    Pathology and Cell Biology, UCL Institute of Ophthalmology, London, United Kingdom
    Moorfields Eye Hospital, London, United Kingdom
  • G. A. Limb
    Pathology and Cell Biology, UCL Institute of Ophthalmology, London, United Kingdom
  • Footnotes
    Commercial Relationships  B. Bhatia, None; S. Singhal, None; J.S. Ellis, None; P.T. Khaw, None; G.A. Limb, None.
  • Footnotes
    Support  Medical Research Council UK, Helen Hamlyn Trust, The Henry Smith Charity
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 5033. doi:https://doi.org/
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      B. Bhatia, S. Singhal, J. S. Ellis, P. T. Khaw, G. A. Limb; Sox2 Is Required for Maintaining the Stem Cell Characteristics of Human Müller Stem Cells From the Adult Retina. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5033. doi: https://doi.org/.

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

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Abstract

Purpose: : Sox2 expression is associated with uncommitted dividing stem and progenitor cells of the developing nervous system and is also expressed in adult neural stem cells. In this study we investigated the role of the transcription factor Sox2 in maintaining the progenitor characteristics of Müller stem cells from the adult human retina.

Methods: : RNA interference was used to investigate the consequences of Sox2 downregulation on the progenitor characteristics of MIO cells. MIO cell lines are spontaneously immortalized Müller cells from the adult human retina that exhibit characteristics of retinal stem cells. Knockdown of Sox2 was achieved using two Sox2 shRNA constructs and one scrambled construct ligated into the pGSU6-GFP vector. Analysis of transfected MIO cells was carried out by RT-PCR, Western blotting and confocal analysis of immunostained cells.

Results: : After knockdown of Sox2 with the shRNA constructs, Müller stem cells adopted a neural morphology and extended long axons with varicosities characteristic of neural cells. Cells transfected with the silencing constructs upregulated markers of differentiating retinal neurons including HuD, Brn3b, Rhodopsin and S-Opsin whilst downregulating neural stem cell markers such as Notch1. Transfection with one of the constructs markedly reduced the proliferating ability of these cells in culture.

Conclusions: : The present observations strongly indicate that Sox2 may play a very important role in maintaining the neural stem cell characteristics of MIO cells. These results also suggest that Sox2 may confer Müller stem cells the potential to regenerate retinal neurons in the adult human eye.

Keywords: Muller cells • differentiation • gene/expression 
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