title: Telomerase immortalization of human corneal endothelial cells yields functional hexagonal monolayers.
abstract: Human corneal endothelial cells (HCEnCs) form a monolayer of hexagonal cells whose main function is to maintain corneal clarity by regulating corneal hydration. HCEnCs are derived from neural crest and are arrested in the post-mitotic state. Thus cell loss due to aging or corneal endothelial disorders leads to corneal edema and blindness-the leading indication for corneal transplantation. Here we show the existence of morphologically distinct subpopulations of HCEnCs that are interspersed among primary cells and exhibit enhanced self-renewal competence and lack of phenotypic signs of cellular senescence. Colonies of these uniform and hexagonal HCEnCs (HCEnC-21) were selectively isolated and demonstrated high proliferative potential that was dependent on endogenous upregulation of telomerase and cyclin D/CDK4. Further transduction of HCEnC-21 with telomerase yielded a highly proliferative corneal endothelial cell line (HCEnT-21T) that was devoid of oncogenic transformation and retained critical corneal endothelial cell characteristics and functionality. This study will significantly impact the fields of corneal cell biology and regenerative medicine.
authors.affiliation: Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: Thore
authors.initials: T
authors.lastname: Schmedt
authors.forename: Yuming
authors.initials: Y
authors.lastname: Chen
authors.forename: Tracy T
authors.initials: TT
authors.lastname: Nguyen
authors.forename: Shimin
authors.initials: S
authors.lastname: Li
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Bonanno
authors.forename: Ula V
authors.initials: UV
authors.lastname: Jurkunas
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D048430:Cell Shape; D016922:Cellular Senescence; D056741:Cyclin D / Q000378:metabolism; D051358:Cyclin-Dependent Kinase 4 / Q000378:metabolism; D004728:Endothelium, Corneal / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D016623:Ion Pumps / Q000378:metabolism; D017136:Ion Transport; D019098:Telomerase / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D015415:Biomarkers; D056741:Cyclin D; D016623:Ion Pumps; D016159:Tumor Suppressor Protein p53; D051358:Cyclin-Dependent Kinase 4; D019098:Telomerase