title: Differentiation of human embryonic stem cells into clinically amenable keratinocytes in an autogenic environment.
abstract: Human embryonic stem cells (hESCs)-derived keratinocytes hold great clinical and research potential. However, the current techniques are hampered by the use of xenogenic components that limits their clinical application. Here we demonstrated an efficient differentiation of H9 hESCs (H9-hESCs) into keratinocytes (H9-Kert) with the minimum use of animal-derived materials. For differentiation, we established two microenvironment systems originated from H9-hESCs (autogenic microenvironment). These autogenic microenvironment systems consist of an autogenic coculture system (ACC) and an autogenic feeder-free system (AFF). In addition, we showed a stage-specific effect of Activin in promoting keratinocyte differentiation from H9-hESCs while repressing the expression of early neural markers in the ACC system. Furthermore, we also explained the effect of Activin in construction of the AFF system made up of extracellular matrix similar to basement membrane extracted from H9-hESC-derived fibroblasts. H9-Kert differentiated in both systems expressed keratinocyte markers at mRNA and protein levels. H9-Kert were also able to undergo terminal differentiation in high Ca(2+) medium. These findings support the transition toward the establishment of an animal-free microenvironment for successful differentiation of hESCs into keratinocytes for potential clinical application.
authors.affiliation: Oral Sciences Disciplines, Faculty of Dentistry, National University of Singapore, Singapore.
authors.forename: Fahad K
authors.initials: FK
authors.lastname: Kidwai
authors.affiliation: Oral Sciences Disciplines, Faculty of Dentistry, National University of Singapore, Singapore; Centre for Stem Cell and Tissue Engineering, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, People's Republic of China.
authors.forename: Hua
authors.initials: H
authors.lastname: Liu
authors.affiliation: Oral Sciences Disciplines, Faculty of Dentistry, National University of Singapore, Singapore.
authors.forename: Wei Seong
authors.initials: WS
authors.lastname: Toh
authors.affiliation: Oral Sciences Disciplines, Faculty of Dentistry, National University of Singapore, Singapore; Plastic Surgery Hospital (Institute), Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Fu
authors.affiliation: Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore.
authors.forename: Doorgesh S
authors.initials: DS
authors.lastname: Jokhun
authors.affiliation: Oral Sciences Disciplines, Faculty of Dentistry, National University of Singapore, Singapore.
authors.forename: Mohammad M
authors.initials: MM
authors.lastname: Movahednia
authors.affiliation: Oral Sciences Disciplines, Faculty of Dentistry, National University of Singapore, Singapore.
authors.forename: Mingming
authors.initials: M
authors.lastname: Li
authors.affiliation: Oral Sciences Disciplines, Faculty of Dentistry, National University of Singapore, Singapore.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zou
authors.affiliation: Department of Oral Pathology, Radiology and Medicine, and Dows, College of Dentistry, The University of Iowa, Iowa City, Iowa, USA.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Squier
authors.affiliation: Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Toan T
authors.initials: TT
authors.lastname: Phan
authors.affiliation: Oral Sciences Disciplines, Faculty of Dentistry, National University of Singapore, Singapore. Electronic address: dencaot@nus.edu.sg.
authors.forename: Tong
authors.initials: T
authors.lastname: Cao
mesh_terms: D028341:Activins / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D055153:Stem Cell Niche; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D017077:Culture Media, Conditioned; D028341:Activins; D014212:Tretinoin; D002118:Calcium