title: Construction of tissue-engineered full-thickness cornea substitute using limbal epithelial cell-like and corneal endothelial cell-like cells derived from human embryonic stem cells.
abstract: The aim of this study was to construct a full-thickness artificial cornea substitute in vitro by coculturing limbal epithelial cell-like (LEC-like) cells and corneal endothelial cell-like (CEC-like) cells derived from human embryonic stem cells (hESCs) on APCM scaffold. A 400 μm thickness, 11 mm diameter APCM lamella containing Bowman's membrane was prepared as the scaffold using trephine and a special apparatus made by ourselves. LEC-like cells and CEC-like cells, derived from hESCs as our previously described, were cocultured on the scaffold using a special insert of 24-well plates that enabled seeding both sides of the scaffold. Three or four layers of epithelium-like cells and a uniform monolayer of CEC-like cells could be observed by H&E staining. The thickness, endothelial cell density, and mechanical properties of the construct were similar to that of native rabbit corneas. Immunofluorescence analysis showed expression of ABCG2 and CK3 in the epithelium-like cell layers and expression of N-cadherin, ZO-1 and Na+/K + ATPase in the CEC-like cells. The corneal substitutes were well integrated within the host corneas, and the transparency increased gradually in 8-week follow-up after transplantation in the rabbits. These results suggest that the strategy we developed is feasible and effective for construction of tissue-engineered full-thickness cornea substitute with critical properties of native cornea.
authors.affiliation: Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, PR China; The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, PR China.
authors.forename: Canwei
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, PR China.
authors.forename: Liqun
authors.initials: L
authors.lastname: Du
authors.affiliation: Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, PR China.
authors.forename: Peng
authors.initials: P
authors.lastname: Sun
authors.affiliation: Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, PR China.
authors.forename: Lin
authors.initials: L
authors.lastname: Shen
authors.affiliation: Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, PR China.
authors.forename: Kunpeng
authors.initials: K
authors.lastname: Pang
authors.affiliation: Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, PR China. Electronic address: xywu8868@163.com.
authors.forename: Xinyi
authors.initials: X
authors.lastname: Wu
mesh_terms: D000818:Animals; D021522:Bioartificial Organs; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000295:instrumentation / Q000379:methods; D003315:Cornea / Q000166:cytology* / Q000254:growth & development*; D016039:Corneal Transplantation / Q000379:methods; D004728:Endothelium, Corneal / Q000166:cytology* / Q000254:growth & development; D019573:Epithelium, Corneal / Q000166:cytology* / Q000254:growth & development; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D006801:Humans; D011817:Rabbits; D013234:Stem Cells; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods*; D054457:Tissue Scaffolds
keywords: Acellular porcine cornea; Corneal endothelium; Corneal epithelium; Human embryonic stem cell; Penetrating keratoplasty; Tissue-engineered cornea