| A reconstituted telomerase-immortalized human corneal epithelium in vivo: a pilot study. | |
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MedLine Citation:
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PMID: 21780919 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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PURPOSE: Telomerase-immortalized human corneal epithelial cells have been reported to stratify and differentiate in vitro similar to native tissue. The purpose of this study was to assess the ability of a telomerase-immortalized human corneal epithelial cell line to generate a full thickness epithelium in vivo in athymic mice. METHODS: Telomerized corneal epithelial cells were transduced with a retroviral vector encoding the herpes simplex thymidine kinase gene. Efficacy of the thymidine kinase suicide gene was confirmed using a live/dead assay. The epithelium was mechanically removed from athymic nude mice and remaining cells were treated with mitomycin C to prevent re-epithelialization. Telomerized corneal epithelial cells were seeded onto the denuded cornea and allowed to adhere for 4 and 24 hours. Cellular attachment was assessed using a fluorescent cell tracker. Stratification and differentiation were assessed after 7 days using phalloidin and a mouse monoclonal antibody to K3. RESULTS: Telomerized corneal epithelial cells were visualized across the denuded stromal surface at 4 and 24 hours, with multi-layering evident at the latter time point. No epithelium was present in the non-treated eye. After 7 days post-transplantation cells stratified into a multilayered epithelium, with positive K3 expression in basal and suprabasal cells. Treatment with ganciclovir induced significant loss of viability in vitro. CONCLUSIONS: The findings in this pilot study demonstrate that telomerized corneal epithelial cells possess the capacity to reconstitute a stratified corneal epithelium in vivo. The introduction of thymidine kinase allowed for the successful induction of cell death in proliferating cells in vitro. Collectively, these data suggest that a telomerase-immortalized corneal epithelial cell line transduced with thymidine kinase represents a potential model for studying differentiation and epithelial-niche interactions in vivo with potential applications in tissue engineering. |
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Authors:
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Danielle M Robertson; Jerry P Kalangara; Rebeccah B Baucom; W Matthew Petroll; H Dwight Cavanagh |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Current eye research Volume: 36 ISSN: 1460-2202 ISO Abbreviation: Curr. Eye Res. Publication Date: 2011 Aug |
Date Detail:
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Created Date: 2011-07-25 Completed Date: 2011-11-01 Revised Date: 2013-03-08 |
Medline Journal Info:
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Nlm Unique ID: 8104312 Medline TA: Curr Eye Res Country: England |
Other Details:
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Languages: eng Pagination: 706-12 Citation Subset: IM |
Affiliation:
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Department of Ophthalmology, The University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390-9057, USA. Danielle.Robertson@utsouthwestern.edu |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Adhesion Cell Death / drug effects Cell Differentiation / physiology Cell Proliferation Cell Transplantation Cells, Cultured Corneal Diseases / surgery* Debridement Disease Models, Animal Epithelium, Corneal / enzymology, transplantation* Female Fluorescent Antibody Technique, Indirect Ganciclovir / toxicity Genes, Transgenic, Suicide / physiology Humans Mice Mice, Inbred BALB C Mice, Nude Pilot Projects Telomerase / physiology* Thymidine Kinase / genetics |
| Grant Support | |
ID/Acronym/Agency:
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EY020799/EY/NEI NIH HHS; R01 EY018219/EY/NEI NIH HHS; R01 EY018219-03/EY/NEI NIH HHS; R24 EY016664-05/EY/NEI NIH HHS |
| Chemical | |
Reg. No./Substance:
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82410-32-0/Ganciclovir; EC 2.7.1.21/Thymidine Kinase; EC 2.7.7.49/Telomerase |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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