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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)
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.
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.
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.
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.
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