title: Regeneration of skin and cornea by tissue engineering.
abstract: Progress in tissue engineering has led to the development of technologies allowing the reconstruction of autologous tissues from the patient's own cells. Thus, tissue-engineered epithelial substitutes produced from cultured skin epithelial cells undergo long-term regeneration after grafting, indicating that functional stem cells were preserved during culture and following grafting. However, these cultured epithelial sheets reconstruct only the upper layer of the skin and lack the mechanical properties associated to the connective tissue of the dermis. We have designed a reconstructed skin entirely made from human cutaneous cells comprising both the dermis and the epidermis, as well as a well-organized basement membrane by a method named the self-assembly approach. In this chapter, protocols to generate reconstructed skin and corneal epithelium suitable for grafting are described in details. The methods include extraction and culture of human skin keratinocytes, human skin fibroblasts as well as rabbit and human corneal epithelial cells, and a complete description of the skin reconstructed by the self-assembly approach and of corneal epithelium reconstructed over a fibrin gel.
authors.affiliation: Experimental Organogenesis Laboratory/LOEX and Department of Surgery and Ophthalmology, Laval University, Sainte-Foy, Quebec, Canada.
authors.forename: Danielle
authors.initials: D
authors.lastname: Larouche
authors.forename: Claudie
authors.initials: C
authors.lastname: Paquet
authors.forename: Julie
authors.initials: J
authors.lastname: Fradette
authors.forename: Patrick
authors.initials: P
authors.lastname: Carrier
authors.forename: François A
authors.initials: FA
authors.lastname: Auger
authors.forename: Lucie
authors.initials: L
authors.lastname: Germain
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003315:Cornea / Q000502:physiology*; D003470:Culture Media; D004847:Epithelial Cells / Q000166:cytology; D005337:Fibrin / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology; D005782:Gels; D006801:Humans; D015603:Keratinocytes / Q000166:cytology; D011817:Rabbits; D012038:Regeneration; D012867:Skin / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D003470:Culture Media; D005782:Gels; D005337:Fibrin