title: Development of functional human oral mucosal epithelial stem/progenitor cell sheets using a feeder-free and serum-free culture system for ocular surface reconstruction.
abstract: Ocular surface reconstruction (OSR) using tissue-engineered cultivated oral mucosal epithelial cell sheets (COMECS) is a promising newly developed treatment for patients with severe ocular surface disease. Until now, this technique has used exogenic and undefined components such as mouse-derived 3T3 feeder cells and fetal bovine serum. To minimize associated risks of zoonotic infection or transmission of unknown pathogens and so establish a safe and effective protocol for the next generation of treatment modality, we developed a novel technique for the COMECS protocol, using a feeder-free and serum-free (FFSF) culture system. Following this new protocol, COMECS exhibited 4-5 layers of well-stratified and differentiated cells, and we successfully produced functional COMECS that included holoclone-type stem cells. Immunohistochemistry confirmed the presence of markers for cell junction (ZO1, Desmoplakin), basement membrane assembly (Collagen 7, Laminin 5), differentiation (K13, K3), proliferation (Ki67) and stem/progenitor cells (p75) in the FFSF COMECS. When transplanted to the ocular surfaces of rabbits, the tissue survived for up to 2 weeks. This study represents a first step toward assessing the development of functional FFSF COMECS for safe and ideal OSR.
authors.affiliation: Department of Frontier Medical Sciences and Technology for Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto 602-0841, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Nakamura
authors.affiliation: Department of Ophthalmology, University of Tokyo Graduate School of Medicine, Tokyo 113-8655, Japan.
authors.forename: Seiichi
authors.initials: S
authors.lastname: Yokoo
authors.affiliation: Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, UK.
authors.forename: Adams J
authors.initials: AJ
authors.lastname: Bentley
authors.affiliation: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto 602-0841, Japan.
authors.forename: Maho
authors.initials: M
authors.lastname: Nagata
authors.affiliation: Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, UK.
authors.forename: Nigel J
authors.initials: NJ
authors.lastname: Fullwood
authors.affiliation: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto 602-0841, Japan.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Inatomi
authors.affiliation: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto 602-0841, Japan.
authors.forename: Chie
authors.initials: C
authors.lastname: Sotozono
authors.affiliation: Department of Ophthalmology, University of Tokyo Graduate School of Medicine, Tokyo 113-8655, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Yamagami
authors.affiliation: Department of Frontier Medical Sciences and Technology for Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto 602-0841, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kinoshita
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D003315:Cornea / Q000166:cytology / Q000378:metabolism; D003316:Corneal Diseases / Q000378:metabolism / Q000628:therapy; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D009061:Mouth Mucosa / Q000166:cytology / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D016895:Culture Media, Serum-Free