April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Substrate Elastic Modulus Influences Retinal Progenitor mRNA Expression in Müller Cells
Author Affiliations & Notes
  • P. K. Lam
    Biology and Biochemistry, Physics and Astronomy,
    The University of Houston, Houston, Texas
  • J. T. Davis
    Physics, Institute for Engineering and Medicine,
    The University of Houston, Houston, Texas
  • Q. Wen
    Biology and Biochemistry, Physics and Astronomy,
    University of Pennsylvania, Philadelphia, Pennsylvania
  • D. C. Otteson
    Biology and Biochemistry, Physics and Astronomy,
    Optometry and Vision Science,
    The University of Houston, Houston, Texas
  • P. A. Janmey
    Physics, Institute for Engineering and Medicine,
    University of Pennsylvania, Philadelphia, Pennsylvania
  • W. J. Foster
    Physics, Institute for Engineering and Medicine,
    The University of Houston, Houston, Texas
    Ophthalmology, Weill-Cornell Medical College, Houston, Texas
  • Footnotes
    Commercial Relationships  P.K. Lam, None; J.T. Davis, None; Q. Wen, None; D.C. Otteson, None; P.A. Janmey, None; W.J. Foster, None.
  • Footnotes
    Support  NIH/NEI/NIBIB EY017112, NIH/NEI EY007551, National Academies Keck Futures Initiative Smart Prosthetics Grant
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 429. doi:
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      P. K. Lam, J. T. Davis, Q. Wen, D. C. Otteson, P. A. Janmey, W. J. Foster; Substrate Elastic Modulus Influences Retinal Progenitor mRNA Expression in Müller Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):429.

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

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Abstract

Purpose: : Tissue elasticity has been increasingly recognized to be a significant factor in cell behavior. Müller cells have demonstrated many characteristics of retinal progenitor cells. To better understand the mechanosensitivity of Müller cells and its possible significance in the development of techniques for retinal regeneration, we examined the gene expression of neurodevelopmental and extracellular matrix genes in Müller cells when cultured on substrates of varying elastic modulus.

Methods: : Conditionally immortalized mouse Müller cells (ImM10) were cultured on laminin-coated polyacrylamide substrates with stiffness of 500 Pa, 1000 Pa, and 5000 Pa, with laminin-coated glass as a control. After 21 days in culture, total RNA was extracted, reverse transcribed, and gene expression analyzed using quantitative real time PCR in triplicate on the genes of interest.

Results: : Of the genes studied, those that demonstrated a continuous increase or decrease in expression of mRNA as a function of the substrate elastic modulus were further examined. We found that the neural progenitor markers Nestin (4-fold change, R2=0.71) and Pax6 (2.3-fold change, R2=0.91), as well as the glial genes Vimentin (1.8-fold change, R2=0.97) and Gfap (2.7-fold change, R2=0.61) are upregulated in Müller cells grown on soft substrates.

Conclusions: : Change in the stiffness of the substrate is associated with changes in expression of some genes that are known neurodevelopmental markers as well as in expression of genes characteristic of both neural and glial development in Müller cells. In addition to the use of growth factors, modification of substrate elastic modulus may facilitate the isolation and controlled differentiation of retinal precursor cells. In therapeutic applications of retinal precursors and stem cells, the local tissue elastic modulus may provide an additional mechanism to control final cell fate.

Keywords: Muller cells • extracellular matrix • retinal culture 
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