title: Comparative proteomics reveals human pluripotent stem cell-derived limbal epithelial stem cells are similar to native ocular surface epithelial cells.
abstract: Limbal epithelial stem cells (LESCs) are tissue-specific stem cells responsible for renewing the corneal epithelium. Acute trauma or chronic disease affecting LESCs may disrupt corneal epithelial renewal, causing vision threatening and painful ocular surface disorders, collectively referred to as LESC deficiency (LESCD). These disorders cannot be treated with traditional corneal transplantation and therefore alternative cell sources for successful cell-based therapy are needed. LESCs derived from human pluripotent stem cells (hPSCs) are a prospective source for ocular surface reconstruction, yet critical evaluation of these cells is crucial before considering clinical applications. In order to quantitatively evaluate hPSC-derived LESCs, we compared protein expression in native human corneal cells to that in hPSC-derived LESCs using isobaric tag for relative and absolute quantitation (iTRAQ) technology. We identified 860 unique proteins present in all samples, including proteins involved in cell cycling, proliferation, differentiation and apoptosis, various LESC niche components, and limbal and corneal epithelial markers. Protein expression profiles were nearly identical in LESCs derived from two different hPSC lines, indicating that the differentiation protocol is reproducible, yielding homogeneous cell populations. Their protein expression profile suggests that hPSC-derived LESCs are similar to the human ocular surface epithelial cells, and possess LESC-like characteristics.
authors.affiliation: BioMediTech, University of Tampere, Finland.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Mikhailova
authors.affiliation: Department of Ophthalmology, School of Medicine, University of Tampere, Finland.
authors.forename: Antti
authors.initials: A
authors.lastname: Jylhä
authors.affiliation: BioMediTech, University of Tampere, Finland.
authors.forename: Jochen
authors.initials: J
authors.lastname: Rieck
authors.affiliation: Department of Ophthalmology, School of Medicine, University of Tampere, Finland.
authors.forename: Janika
authors.initials: J
authors.lastname: Nättinen
authors.affiliation: BioMediTech, University of Tampere, Finland.
authors.forename: Tanja
authors.initials: T
authors.lastname: Ilmarinen
authors.affiliation: Stem Cells and Eye Research Laboratory, Department of Ophthalmology, Faculty of Medicine, University of Szeged, Hungary.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Veréb
authors.affiliation: Department of Ophthalmology, School of Medicine, University of Tampere, Finland.
authors.forename: Ulla
authors.initials: U
authors.lastname: Aapola
authors.affiliation: Department of Ophthalmology, School of Medicine, University of Tampere, Finland.|Singapore Eye Research Institute and School of Medicine, Singapore.
authors.forename: Roger
authors.initials: R
authors.lastname: Beuerman
authors.affiliation: Stem Cells and Eye Research Laboratory, Department of Ophthalmology, Faculty of Medicine, University of Szeged, Hungary.
authors.forename: Goran
authors.initials: G
authors.lastname: Petrovski
authors.affiliation: Department of Ophthalmology, School of Medicine, University of Tampere, Finland.|Tampere University Hospital Eye Center, University of Tampere, Finland.
authors.forename: Hannu
authors.initials: H
authors.lastname: Uusitalo
authors.affiliation: BioMediTech, University of Tampere, Finland.
authors.forename: Heli
authors.initials: H
authors.lastname: Skottman
mesh_terms: D000368:Aged; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D004847:Epithelial Cells / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000166:cytology; D000066449:Human Embryonic Stem Cells / Q000378:metabolism; D006801:Humans; D008297:Male; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D055153:Stem Cell Niche
chemical_list: D051017:Apoptosis Regulatory Proteins; D015415:Biomarkers; D020543:Proteome