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PS Carrier, A Deschambeault, M Talbot, SL Guerin, FA Auger, L Germain; Expression Of Extracellular Matrix Components In Reconstructed Human Cornea Produced By Tissue Engineering . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4166.
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Purpose:We have reported a new tissue engineering method for the production of fully human reconstructed tissues, from mesenchymal and epithelial cells, without the addition of exogenous extracellular matrix proteins. This auto-assembly approach was used to produce cornea (from human keratocytes and corneal epithelial cells) and skin (from fibroblasts and skin epithelial cells). The expression of extracellular matrix components (collagens, fibronectin and laminin) were studied in these reconstructed cornea and skin. The results were compared with the in situ expression of the same components in the cornea (including the limbus) and skin. Methods:Reconstructed human cornea and skin were produced by culturing keratocytes or fibroblasts in the presence of ascorbic acid to produce sheets on which epithelial cells were then seeded and cultured. Histological (Masson's trichrome staining) and immunohistological (type I, II, III, IV, V, VI, VII, XII, XIV collagen, laminin and fibronectin) studies were performed on the reconstructed tissues and frozen biopsies from normal human cornea and skin. Results:Reconstructed corneas presented a different histological and extracellular matrix components' expression pattern compared to reconstructed skin. These patterns resembled that of their respective native tissues. We have also observed that the expression pattern of these extracellular matrix proteins was different in the limbus compared to the center of the cornea. Conclusion:Our results provide evidence that we are able to produce in vitro reconstructed human cornea and skin exhibiting distinct structural and histological properties corresponding to their native tissues. Moreover, our observations show a different pattern of expression of extracellular matrix components in the cornea, the limbus and the skin.
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