Document Detail


Culture, immortalization, and characterization of human meibomian gland epithelial cells.
MedLine Citation:
PMID:  20335607     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: Meibomian gland epithelial cells are essential in maintaining the health and integrity of the ocular surface. However, very little is known about their physiological regulation. In this study, the cellular control mechanisms were explored, first to establish a defined culture system for the maintenance of primary epithelial cells from human meibomian glands and, second, to immortalize these cells, thereby developing a preclinical model that could be used to identify factors that regulate cell activity.
METHODS: Human meibomian glands were removed from lid segments after surgery, enzymatically digested, and dissociated. Isolated epithelial cells were cultured in media with or without serum and/or 3T3 feeder layers. To attempt immortalization, the cells were exposed to retroviral human telomerase reverse transcriptase (hTERT) and/or SV40 large T antigen cDNA vectors, and antibiotic-resistant cells were selected, expanded, and subcultured. Analyses for possible biomarkers, cell proliferation and differentiation, lipid-related enzyme gene expression, and the cellular response to androgen were performed with biochemical, histologic, and molecular biological techniques.
RESULTS: It was possible to isolate viable human meibomian gland epithelial cells and to culture them in serum-free medium. These cells proliferated, survived through at least the fifth passage, and contained neutral lipids. Infection with hTERT immortalized these cells, which accumulated neutral lipids during differentiation, expressed multiple genes for lipogenic enzymes, responded to androgen, and continued to proliferate.
CONCLUSIONS: The results show that human meibomian gland epithelial cells may be isolated, cultured, and immortalized.
Authors:
Shaohui Liu; Mark P Hatton; Payal Khandelwal; David A Sullivan
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2010-03-24
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  51     ISSN:  1552-5783     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2010 Aug 
Date Detail:
Created Date:  2010-07-23     Completed Date:  2010-08-20     Revised Date:  2011-07-22    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3993-4005     Citation Subset:  IM    
Affiliation:
Schepens Eye Research Institute, Harvard Medical School, Boston, Massachusetts 02114, USA.
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MeSH Terms
Descriptor/Qualifier:
Adult
Aged
Aged, 80 and over
Antigens, Polyomavirus Transforming / genetics,  metabolism
Cell Culture Techniques
Cell Division
Cell Line
Cell Proliferation
Epithelial Cells / cytology*,  metabolism
Female
Fluorescent Antibody Technique, Indirect
Genetic Vectors
Humans
Karyotyping
Lipid Metabolism
Male
Meibomian Glands / cytology*,  metabolism
Middle Aged
Reverse Transcriptase Polymerase Chain Reaction
Telomerase / genetics,  metabolism
Grant Support
ID/Acronym/Agency:
EY05612/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/Antigens, Polyomavirus Transforming; EC 2.7.7.49/TERT protein, human; EC 2.7.7.49/Telomerase
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