April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Optimisation of Cellular Scaffolds for Neural Retinal Cell Replacement Using Human Müller Stem Cells
Author Affiliations & Notes
  • L. M. James
    ORBIT, Ocular Biology and Therapeutics,
    Institue of Ophthalmology UCL, London, United Kingdom
  • S. Singhal
    ORBIT, Ocular Biology and Therapeutics,
    UCL Institute of Ophthalmology, London, United Kingdom
  • H. Jayaram
    Ocular Biology and Therapeutics, Ocular Biology & Therapeutics,
    UCL Institute of Ophthalmology, London, United Kingdom
  • P. T. Khaw
    Director, Research & Development, Moorfields Eye Hospital & UCL Inst Ophth, London, United Kingdom
  • G. A. Limb
    Ocular Biology and Therapeutics, Ocular Biology & Therapeutics,
    Institue of Ophthalmology UCL, London, United Kingdom
  • Footnotes
    Commercial Relationships  L.M. James, None; S. Singhal, None; H. Jayaram, None; P.T. Khaw, None; G.A. Limb, None.
  • Footnotes
    Support  Biomedical Research centre for Ophthalmology Moorfield's eye hospital, MRC
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 3146. doi:
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    • Get Citation

      L. M. James, S. Singhal, H. Jayaram, P. T. Khaw, G. A. Limb; Optimisation of Cellular Scaffolds for Neural Retinal Cell Replacement Using Human Müller Stem Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3146.

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

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Abstract

Purpose: : Adult human Müller stem cells can be made to differentiate into retinal neurons in vitro. These cells may be potentially used for retinal transplantation to replace neurons damaged as a result of retinal degeneration. This study aimed to investigate the use of Collagen type I as a source of biomaterial to build cellular scaffolds using Müller stem cells.

Methods: : Nanofibre mats of Collagen type I were used to culture Müller stem cells in the presence of the Notch1 inhibitor DAPT (N-[N-(3, 5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester) and FGF2 for 1 week. Cellular attachment and neural morphology were examined by phase microscopy and scanning electron microscopy. Expression of the retinal ganglion cell markers HuD and Islet-1 was examined by confocal microscopy.

Results: : Cells cultured for one week on nanofibre mats of Collagen type-I in the presence of DAPT and FGF2 displayed long processes characteristic of neural morphology. These cellular processes were more pronounced when cells were cultured on thin Collagen nanofibre mats than when cultured on thick mats. Cells cultured on these scaffolds also showed immunostaining for the retinal ganglion cell markers HuD and Islet-1.

Conclusions: : Nanofibre mats of Collagen type I provide the mechanical and physical properties necessary for cell delivery into the degenerated retina. These mats support adhesion and spreading of human Müller stem cells as well as neurite formation upon differentiation into the ganglion cell phenotype. Further studies will explore the practical application of these scaffolds in cell based therapies to repair damaged retina.

Keywords: Muller cells • ganglion cells • transplantation 
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