Abstract
Tissue engineering is a flourishing field of regenerative medicine that allows the reconstruction of various tissues of our body, including the cornea. In addition to addressing the growing need for organ transplants, such tissue-engineered substitutes may also serve as good in vitro models for fundamental and preclinical studies. Recent progress in the field of corneal tissue engineering has led to the development of new technologies allowing the reconstruction of a human bi-lamellar cornea. One unique feature of this model is the complete absence of exogenous material. Indeed, these human corneal equivalents are exclusively composed of untransformed human corneal fibroblasts (hCFs) entangled in their own extracellular matrix, as well as untransformed human corneal epithelial cells (hCECs), both of which isolated from donor corneas. The reconstructed human bi-lamellar cornea thereby exhibits a well-organized stroma as well as a well-differentiated epithelium. This chapter describes the methods used for the isolation and culture of hCFs, the production and assembly of hCFs stromal sheets, the seeding of hCECs, and the maturation of the tissue-engineered cornea.
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Acknowledgements
The authors would like to thank current and former members of the LOEX and CUO-Recherche laboratories who contributed to develop and improve the foregoing protocols. This work was supported by the Canadian Institutes for Health Research (CIHR) grant MOP-12087 and FDN-143213 (L.G.), the Fondation des Pompiers du Québec pour les Grands Brûlés (FPQGB), the Fonds de Recherche du Québec-Santé (FRQS), and the Réseau de thérapie cellulaire, tissulaire et génique du Québec -ThéCell (a thematic network supported by the FRQS). The Banque d’yeux Nationale is partly supported by the Réseau de Recherche en Santé de la Vision from the FRQS. P.D. and C.C. were supported by studentships from the FRQS. L.G. is the recipient of a Tier 1 Canadian Research Chair on Stem Cells and Tissue Engineering and a Research Chair on Tissue-Engineered Organs and Translational Medicine of the Fondation de l’Université Laval.
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Le-Bel, G., Desjardins, P., Couture, C., Germain, L., Guérin, S.L. (2020). The Self-assembly Approach as a Tool for the Tissue Engineering of a Bi-lamellar Human Cornea. In: Ahearne, M. (eds) Corneal Regeneration. Methods in Molecular Biology, vol 2145. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0599-8_8
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DOI: https://doi.org/10.1007/978-1-0716-0599-8_8
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