Document Detail


A reconstituted telomerase-immortalized human corneal epithelium in vivo: a pilot study.
MedLine Citation:
PMID:  21780919     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Danielle M Robertson; Jerry P Kalangara; Rebeccah B Baucom; W Matthew Petroll; H Dwight Cavanagh
Related Documents :
21959449 - Isolation of epithelial cells in the developing primary lip and palate.
18287499 - Complementary modulation of somatic inhibition by opioids and cannabinoids.
21590129 - Amifostine (wr-2721, ethyol(tm)) can protect human endothelial but not tumor cells from...
22244889 - Crkl plays a pivotal role in tumorigenesis of head and neck squamous cell carcinoma thr...
21696969 - 18β-glycyrrhetinic acid derivatives induced mitochondrial-mediated apoptosis through r...
21630659 - Anticancer activities of thelephantin o and vialinin a isolated from thelephora auranti...
15782099 - Similarities of k+atp channel expression and ca2+ changes in pancreatic beta cells and ...
11472759 - Surface characteristics of foreign targets that elicit an encapsulation response by the...
10455499 - A single-use luciferase-based mercury biosensor using escherichia coli hb101 immobilize...
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Current eye research     Volume:  36     ISSN:  1460-2202     ISO Abbreviation:  Curr. Eye Res.     Publication Date:  2011 Aug 
Date Detail:
Created Date:  2011-07-25     Completed Date:  2011-11-01     Revised Date:  2013-03-08    
Medline Journal Info:
Nlm Unique ID:  8104312     Medline TA:  Curr Eye Res     Country:  England    
Other Details:
Languages:  eng     Pagination:  706-12     Citation Subset:  IM    
Affiliation:
Department of Ophthalmology, The University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390-9057, USA. Danielle.Robertson@utsouthwestern.edu
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
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:
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:
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


Previous Document:  Effect of sub-atmospheric oxygen on the culture of rabbit limbal epithelial cells.
Next Document:  Antioxidant status and oxidative stress in primary open angle glaucoma and pseudoexfoliative glaucom...