title: Characterization of biomaterial-free cell sheets cultured from human oral mucosal epithelial cells.
abstract: The purpose of this study was to report the characteristics of biomaterial-free sheets cultured from human oral mucosal epithelial cells without fibrin support, in vitro and after transplantation to limbal-deficient models. Human oral mucosal epithelial cells and limbal epithelial cells were cultured for 2 weeks, and the colony-forming efficiency (CFE) rates were compared. Markers of stem cells (p63), cell proliferation (Ki-67) and epithelial differentiation (cytokeratin; K1, K3, K4, K13) were observed in colonies and in biomaterial-free sheets. Biomaterial-free sheets which had been detached with 1% dispase or biomaterial-free sheets generated by fibrin support were transplanted to 12 limbal-deficient rabbit models. In vitro cell viability, in vivo stability and cytokeratin characteristics of biomaterial-free sheets were compared with those of sheets formed by fibrin-coated culture 1 week after transplantation. Mean CFE rate was significantly higher in human oral mucosal epithelial cells (44.8%) than in human limbal epithelial cells(17.7%). K3 and K4 were well expressed in both colonies and sheets. Biomaterial-free sheets had two to six layers of stratified cells and showed an average of 79.8% viable cells in the sheets after detachment. Cytokeratin expressions of biomaterial-free sheets were comparable to those of sheets cultured by fibrin support, in limbal-deficient models. Both p63 and Ki-67 were well expressed in colonies, isolated sheets and sheets transplanted to limbal-deficient models. Our results suggest that biomaterial-free sheets cultured from human oral mucosal epithelial cells without fibrin support can be an alternative option for cell therapy in use for the treatment of limbal-deficient diseases. Copyright © 2014 John Wiley & Sons, Ltd.
authors.affiliation: Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea.|Laboraory of Ocular Regenerative Medicine and Immunology, Seoul Artificial Eye Centre, Seoul National University Hospital Biomedical Research Institute, Seoul, Korea.
authors.forename: Dong Won
authors.initials: DW
authors.lastname: Hyun
authors.affiliation: Cutigen Research Institute, Tego Science Inc, Seoul, Korea.
authors.forename: Yun Hee
authors.initials: YH
authors.lastname: Kim
authors.affiliation: Laboraory of Ocular Regenerative Medicine and Immunology, Seoul Artificial Eye Centre, Seoul National University Hospital Biomedical Research Institute, Seoul, Korea.
authors.forename: Ah Young
authors.initials: AY
authors.lastname: Koh
authors.affiliation: Laboraory of Ocular Regenerative Medicine and Immunology, Seoul Artificial Eye Centre, Seoul National University Hospital Biomedical Research Institute, Seoul, Korea.
authors.forename: Hyun Ju
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea.|Laboraory of Ocular Regenerative Medicine and Immunology, Seoul Artificial Eye Centre, Seoul National University Hospital Biomedical Research Institute, Seoul, Korea.
authors.forename: Won Ryang
authors.initials: WR
authors.lastname: Wee
authors.affiliation: Cutigen Research Institute, Tego Science Inc, Seoul, Korea.
authors.forename: Saewha
authors.initials: S
authors.lastname: Jeon
authors.affiliation: Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea.|Laboraory of Ocular Regenerative Medicine and Immunology, Seoul Artificial Eye Centre, Seoul National University Hospital Biomedical Research Institute, Seoul, Korea.
authors.forename: Mee Kum
authors.initials: MK
authors.lastname: Kim
mesh_terms: D001672:Biocompatible Materials / Q000494:pharmacology*; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D020397:Goblet Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007633:Keratins / Q000378:metabolism; D009061:Mouth Mucosa / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
chemical_list: D001672:Biocompatible Materials; D015415:Biomarkers; D007633:Keratins
keywords: biomaterial-free sheet; colony-forming efficiency (CFE); cornea; limbal stem cell deficiency disease (LSCD); ocular surface; oral epithelial cell sheet