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C.J. Giasson, P. Carrier, L. Germain; Collagen-based Corneal Equivalents Made with Human Cells as a Model to Study Cell-cell Contact . Invest. Ophthalmol. Vis. Sci. 2003;44(13):927.
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Purpose: Observe the structure of corneal equivalents and test for the presence of cadherin, the adhering junction protein, in epithelial and endothelial layers bordering a reconstructed stroma. Methods: Cells came from eyes unsuitable for transplantation from the National eyebank. Corneal fibroblasts as well as epithelial and endothelial cells were harvested and grown separately. At confluence, a stromal equivalent was constructed by seeding first fibroblasts into a collagen gel, then, epithelial and endothelial cells on each side of this stroma. Specimens were stained with Masson's trichrome or dissected in order to lyse separately epithelial and endothelial cells. Cell lysates proteins separated by SDS-PAGE were transferred to membranes and expose to pan-cadherin antibody. Results: A middle blue stroma, containing fibroblastic cells, was lined by a monolayered endothelium. On the other side, 3 or 4 layers of epithelial cells were present; basal cells were more cuboidal and superficial cells, flatter. Cadherin was detected in both lysates. Conclusions: The structure of this collagen-based corneal equivalent simulates the native cornea. Cells from the epithelial and endothelial layers express cadherins, suggesting the presence of cell-cell contacts.
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