title: Establishment of a novel in vitro model of stratified epithelial wound healing with barrier function.
abstract: The repair of wounds through collective movement of epithelial cells is a fundamental process in multicellular organisms. In stratified epithelia such as the cornea and skin, healing occurs in three steps that include a latent, migratory, and reconstruction phases. Several simple and inexpensive assays have been developed to study the biology of cell migration in vitro. However, these assays are mostly based on monolayer systems that fail to reproduce the differentiation processes associated to multilayered systems. Here, we describe a straightforward in vitro wound assay to evaluate the healing and restoration of barrier function in stratified human corneal epithelial cells. In this assay, circular punch injuries lead to the collective migration of the epithelium as coherent sheets. The closure of the wound was associated with the restoration of the transcellular barrier and the re-establishment of apical intercellular junctions. Altogether, this new model of wound healing provides an important research tool to study the mechanisms leading to barrier function in stratified epithelia and may facilitate the development of future therapeutic applications.
authors.affiliation: Schepens Eye Research Institute and Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Miguel
authors.initials: M
authors.lastname: Gonzalez-Andrades
authors.affiliation: Changing Places Research Group, Media Lab, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Luis
authors.initials: L
authors.lastname: Alonso-Pastor
authors.affiliation: Schepens Eye Research Institute and Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Mauris
authors.affiliation: Schepens Eye Research Institute and Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Andrea
authors.initials: A
authors.lastname: Cruzat
authors.affiliation: Schepens Eye Research Institute and Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Claes H
authors.initials: CH
authors.lastname: Dohlman
authors.affiliation: Schepens Eye Research Institute and Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Pablo
authors.initials: P
authors.lastname: Argüeso
mesh_terms: D002465:Cell Movement; D004847:Epithelial Cells / Q000378:metabolism; D004848:Epithelium / Q000473:pathology* / Q000502:physiology*; D019573:Epithelium, Corneal / Q000473:pathology / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D012038:Regeneration; D014945:Wound Healing