title: Differentiation of human embryonic stem cells into corneal epithelial-like cells by in vitro replication of the corneal epithelial stem cell niche.
abstract: Human embryonic stem cells (hESCs) are pluripotent cells capable of differentiating into any cell type of the body. It has long been known that the adult stem cell niche is vital for the maintenance of adult stem cells. The cornea at the front of the eye is covered by a stratified epithelium that is renewed by stem cells located at its periphery in a region known as the limbus. These so-called limbal stem cells are maintained by factors within the limbal microenvironment, including collagen IV in basement membrane and limbal fibroblasts in the stroma. Because this niche is very specific to the stem cells (rather than to the more differentiated cells) of the corneal epithelium, it was hypothesized that replication of these factors in vitro would result in hESC differentiation into corneal epithelial-like cells. Indeed, here we show that culturing of hESC on collagen IV using medium conditioned by the limbal fibroblasts results in the loss of pluripotency and differentiation into epithelial-like cells. Further differentiation results in the formation of terminally differentiated epithelial-like cells not only of the cornea but also of skin. Scanning electron microscopy shows that some differences exist between hESC-derived and adult limbal epithelial-like cells, necessitating further investigation using in vivo animal models of limbal stem cell deficiency. Such a model of hESC differentiation is useful for understanding the early events of epithelial lineage specification and to the eventual potential application of epithelium differentiated from hESC for clinical conditions of epithelial stem cell loss. Disclosure of potential conflicts of interest is found at the end of this article.
authors.affiliation: Centre for Stem Cell Biology and Developmental Genetics, University of Newcastle, Newcastle upon Tyne, United Kingdom.
authors.forename: Sajjad
authors.initials: S
authors.lastname: Ahmad
authors.forename: Rebecca
authors.initials: R
authors.lastname: Stewart
authors.forename: Sun
authors.initials: S
authors.lastname: Yung
authors.forename: Sai
authors.initials: S
authors.lastname: Kolli
authors.forename: Lyle
authors.initials: L
authors.lastname: Armstrong
authors.forename: Miodrag
authors.initials: M
authors.lastname: Stojkovic
authors.forename: Francisco
authors.initials: F
authors.lastname: Figueiredo
authors.forename: Majlinda
authors.initials: M
authors.lastname: Lako
mesh_terms: D016475:3T3 Cells; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000648:ultrastructure; D004847:Epithelial Cells / Q000166:cytology*; D019573:Epithelium, Corneal / Q000166:cytology*; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology; D005786:Gene Expression Regulation; D006028:Glycosphingolipids / Q000378:metabolism; D006801:Humans; D007633:Keratins / Q000235:genetics / Q000378:metabolism; D016850:Limbus Corneae / Q000166:cytology; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology; D055746:Stage-Specific Embryonic Antigens; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C121278:CK-34 beta E12; D004268:DNA-Binding Proteins; D006028:Glycosphingolipids; D050814:Octamer Transcription Factor-3; D055746:Stage-Specific Embryonic Antigens; C115051:TP63 protein, human; D015534:Trans-Activators; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C066584:stage-specific embryonic antigen-4; D007633:Keratins