NDLI: Culture, immortalization, and characterization of human meibomian gland epithelial cells.

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Culture, immortalization, and characterization of human meibomian gland epithelial cells.

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Culture, immortalization, and characterization of human meibomian gland epithelial cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Sullivan, David A. Liu, Shaohui Hatton, Mark P. Khandelwal, Payal
Description	Country affiliation: United States Author Affiliation: Liu S ( Schepens Eye Research Institute, Harvard Medical School, Boston, Massachusetts 02114, USA.)
Abstract	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.
ISSN	01460404
e-ISSN	15525783
DOI	10.1167/iovs.09-5108
Journal	Investigative Opthalmology & Visual Science
Issue Number	8
Volume Number	51
Language	English
Publisher	Association for Research in Vision and Ophthalmology
Publisher Date	2010-08-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Epithelial Cells Cytology Meibomian Glands Antigens, Polyomavirus Transforming Genetics Metabolism Cell Culture Techniques Cell Division Cell Line Cell Proliferation Fluorescent Antibody Technique, Indirect Genetic Vectors Karyotyping Lipid Metabolism Reverse Transcriptase Polymerase Chain Reaction Telomerase Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Discipline Ophthalmology
Content Type	Text
Resource Type	Article
Subject	Ophthalmology Sensory Systems Cellular and Molecular Neuroscience

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