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S. Proulx, J. Uwamaliya, C. Audet, A. Deschambeault, P. Carrier, I. Brunette, L. Germain; Reconstruction of the Corneal Endothelium in the Feline Model. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5728.
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To characterize a reconstructed corneal endothelium bioengineered by seeding previously cultured feline endothelial cells onto denuded Descemet’s membrane of human corneas.
Second-passaged feline endothelial cells were seeded on top of a previously decellularized human cornea. After 2 weeks of culture to allow for attachment and spreading of seeded cells, the reconstructed corneas were stained with alizarin red for endothelial cell count and fixed for histology and scanning (SEM) and transmission (TEM) electron microscopy. Immunofluorescence labelling of of various proteins was also performed.
Alizarin staining showed that the reconstructed feline corneal endothelial mean cell counts were 2289 ± 233 cells/mm2. Histology and TEM showed that the reconstructed endothelium formed a monolayer of tightly-packed cells that were well adhered to Descemet’s membrane. SEM confirmed that the endothelium covered the entire posterior corneal surface and that the cells adopted an endothelial morphology. The reconstructed feline corneal endothelium also expressed K8/18 confirming its endothelial origin.
This study demonstrates the feasibility of reconstructing a highly cellular and healthy corneal endothelium. Potential applications of this model are numerous, including in vitro pharmacological studies. This represents an additional step towards the future development bioengineered corneas.
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