title: Small-molecule induction promotes corneal epithelial cell differentiation from human induced pluripotent stem cells.
abstract: Human induced pluripotent stem cells (hiPSCs) offer unique opportunities for developing novel cell-based therapies and disease modeling. In this study, we developed a directed differentiation method for hiPSCs toward corneal epithelial progenitor cells capable of terminal differentiation toward mature corneal epithelial-like cells. In order to improve the efficiency and reproducibility of our method, we replicated signaling cues active during ocular surface ectoderm development with the help of two small-molecule inhibitors in combination with basic fibroblast growth factor (bFGF) in serum-free and feeder-free conditions. First, small-molecule induction downregulated the expression of pluripotency markers while upregulating several transcription factors essential for normal eye development. Second, protein expression of the corneal epithelial progenitor marker p63 was greatly enhanced, with up to 95% of cells being p63 positive after 5 weeks of differentiation. Third, corneal epithelial-like cells were obtained upon further maturation.
authors: Institute of Biomedical Technology, University of Tampere, Tampere 33520, Finland ; BioMediTech, Tampere 33520, Finland.
authors: Alexandra
authors: A
authors: Mikhailova
authors: Institute of Biomedical Technology, University of Tampere, Tampere 33520, Finland ; BioMediTech, Tampere 33520, Finland.
authors: Tanja
authors: T
authors: Ilmarinen
authors: SILK, Department of Ophthalmology, University of Tampere, Tampere 33520, Finland ; TAUH Eye Center, Tampere University Hospital, Tampere 33520, Finland.
authors: Hannu
authors: H
authors: Uusitalo
authors: Institute of Biomedical Technology, University of Tampere, Tampere 33520, Finland ; BioMediTech, Tampere 33520, Finland.
authors: Heli
authors: H
authors: Skottman
mesh_terms: D052117:Benzodioxoles / Q000494:pharmacology; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D019573:Epithelium, Corneal / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D005123:Eye / Q000196:embryology / Q000378:metabolism; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors; D060449:Wnt Signaling Pathway / Q000187:drug effects
chemical_list: C519132:2-(5-benzo(1,3)dioxol-5-yl-2-tert-butyl-3H-imidazol-4-yl)-6-methylpyridine hydrochloride; D052117:Benzodioxoles; C081876:CKAP4 protein, human; D007093:Imidazoles; D008565:Membrane Proteins; D011725:Pyridines; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D005346:Fibroblast Growth Factors