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

Shaohui Liu, Mark P. Hatton, Payal Khandelwal, David A. Sullivan

Research output: Contribution to journalArticle

54 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)3993-4005
Number of pages13
JournalInvestigative Ophthalmology and Visual Science
Volume51
Issue number8
DOIs
StatePublished - Aug 1 2010
Externally publishedYes

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Meibomian Glands
Epithelial Cells
Lipids
Androgens
Polyomavirus Transforming Antigens
Feeder Cells
Viral Tumor Antigens
Serum-Free Culture Media
Enzymes
Cell Differentiation
Complementary DNA
Cell Culture Techniques
Biomarkers
Maintenance
Cell Proliferation
Anti-Bacterial Agents
Gene Expression
Health
Infection
Serum

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Culture, immortalization, and characterization of human meibomian gland epithelial cells. / Liu, Shaohui; Hatton, Mark P.; Khandelwal, Payal; Sullivan, David A.

In: Investigative Ophthalmology and Visual Science, Vol. 51, No. 8, 01.08.2010, p. 3993-4005.

Research output: Contribution to journalArticle

Liu, Shaohui ; Hatton, Mark P. ; Khandelwal, Payal ; Sullivan, David A. / Culture, immortalization, and characterization of human meibomian gland epithelial cells. In: Investigative Ophthalmology and Visual Science. 2010 ; Vol. 51, No. 8. pp. 3993-4005.
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