Reconstruction of a human cornea by the self-assembly approach of tissue engineering using the three native cell types

Authors: Proulx, StéphanieUwamaliya, Jeanne d'ArcCarrier, PatrickDeschambeault, AlexandreAudet, CarolineGiasson, Claude-J.Guérin, SylvainAuger, François A.Germain, Lucie
Abstract: Purpose: The purpose of this study was to produce and characterize human tissue-engineered corneas reconstructed using all three corneal cell types (epithelial, stromal, and endothelial cells) by the self-assembly approach. Methods: Fibroblasts cultured in medium containing serum and ascorbic acid secreted their own extracellular matrix and formed sheets that were superposed to reconstruct a stromal tissue. Endothelial and epithelial cells were seeded on each side of the reconstructed stroma. After culturing at the air-liquid interface, the engineered corneas were fixed for histology and transmission electron microscopy (TEM). Immunofluorescence labeling of epithelial keratins, basement membrane components, Na+/K+-ATPase α1, and collagen type I was also performed. Results: Epithelial and endothelial cells adhered to the reconstructed stroma. After 10 days at the air-liquid interface, the corneal epithelial cells stratified (4 to 5 cell layers) and differentiated into well defined basal and wing cells that also expressed Na+/K+-ATPase α1 protein, keratin 3/12, and basic keratins. Basal epithelial cells from the reconstructed epithelium formed many hemidesmosomes and secreted a well defined basement membrane rich in laminin V and collagen VII. Endothelial cells formed a monolayer of tightly-packed cells and also expressed the function related protein Na+/K +-ATPase α1. Conclusions: This study demonstrates the feasibility of producing a complete tissue-engineered human cornea, similar to native corneas, using untransformed fibroblasts, epithelial and endothelial cells, without the need for exogenous biomaterial.
Document Type: Article de recherche
Issue Date: 29 October 2010
Open Access Date: 29 January 2018
Document version: VoR
This document was published in: Molecular vision, Vol. 16 (234-236), 2192–2201 (2010)
Éditeur non identifié
Alternative version: 21139684
Collection:Articles publiés dans des revues avec comité de lecture

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