Impact of cell source on human cornea reconstructed by tissue engineering

Authors: Carrier, PatrickDeschambeault, AlexandreAudet, CarolineGiroux-Talbot, MarièveGauvin, RobertGiasson, Claude-J.Auger, François A.Guérin, SylvainGermain, Lucie
Abstract: Purpose: To investigate the effect of the tissue origin of stromal fibroblasts and epithelial cells on reconstructed corneas in vitro. Methods: Four types of constructs were produced by the self-assembly approach using the following combinations of human cells: corneal fibroblasts/corneal epithelial cells, corneal fibroblasts/skin epithelial cells, skin fibroblasts/corneal epithelial cells, skin fibroblasts/skin epithelial cells. Fibroblasts were cultured with ascorbic acid to produce stromal sheets on which epithelial cells were cultured. After 2 weeks at the air-liquid interface, the reconstructed tissues were photographed, absorption spectra were measured, and tissues were fixed for histologic analysis. Cytokine expression in corneal- or skin-fibroblast-conditioned media was determined with the use of protein array membranes. The effect of culturing reconstructed tissues with conditioned media, or media supplemented with a cytokine secreted mainly by corneal fibroblasts, was determined. Results: The tissue source from which epithelial and mesenchymal cells were isolated had a great impact on the macroscopic and histologic features (epithelium thickness and differentiation) and the functional properties (transparency) of the reconstructed tissues. The reconstructed cornea had ultraviolet-absorption characteristics resembling those of native human cornea. The regulation of epithelial differentiation and thickness was mesenchyme-dependent and mediated by diffusible factors. IL-6, which is secreted in greater amounts by corneal fibroblasts than skin fibroblasts, decreased the expression of the differentiation marker DLK in the reconstructed epidermis. Conclusions: The tissue origin of fibroblasts and epithelial cells plays a significant role in the properties of the reconstructed tissues. These human models are promising tools for gaining a thorough understanding of epithelial-stromal interactions and regulation of epithelia homeostasis.
Document Type: Article de recherche
Issue Date: 1 June 2009
Open Access Date: 29 January 2018
Document version: VoR
Permalink: http://hdl.handle.net/20.500.11794/16728
This document was published in: Investigative ophthalmology & visual science, Vol. 50 (6), 2645 (2009)
https://doi.org/10.1167/iovs.08-2001
IOVS
Alternative version: 10.1167/iovs.08-2001
19218610
Collection:Articles publiés dans des revues avec comité de lecture

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