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A. K. Salz, S. Kaempf, S. Johnen, C. Maltusch, R. Lindt, P. Walter, G. Thumann; Tissue Engineering of Cell Monolayers for Subretinal Transplantation. Invest. Ophthalmol. Vis. Sci. 2008;49(13):279. doi: https://doi.org/.
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The transplantation of pigmented cells in the subretinal space is discussed as a possible therapeutic option for age-related macular degeneration. Previous studies have shown that the transplantation of cell suspensions into the subretinal space does not result in the formation of a cell monolayer and thus the recovery of vision. For the recovery of vision it will be critical to transplant cell monolayers at the location of the exposed photoreceptors to reform the normal morphological and functional relationships between photoreceptors and pigment cells.
To establish pre-formed cell monolayers, ARPE-19 cells were cultured on equine collagen type I membranes. Contact angle measurements and SEM were performed to define the surface characteristics of the collagen membranes. Cell morphology, viability, proliferation and phagocytosis of ROS were investigated. Cytochemical analysis for the expression of cytokeratin, actin, RPE65, PEDF, and ZO-1 was used to evaluate differentiated characteristics. In addition, the response of ARPE-19 cells to genetical modification by non-viral transfection was examined.
ARPE-19 cells cultured on collagen membranes adhere, proliferate and form confluent monolayers. The collagen foil demonstrates excellent hydrophilicity with a contact angel of 66°. Surface morphology of the membrane as visualized by SEM appeared to have similar features of Bruch’s membrane. Cells grown as a monolayer on the collagen membranes acquire differentiated properties and show enhanced phagocytosis and are amenable to be transfected with GFP-plasmids with an efficiency of 82%.
Pigment epithelial cells cultured on collagen membranes acquire differentiated properties of in vivo retinal pigment epithelial cells and form complete monolayers that are amenable to be transplanted to the subretinal space.
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