Abstract
PurposeThe aim of this study was to evaluate a human meibomian gland epithelial cell line (HMGEC) as a model for meibomian gland (patho)physiology in vitro.MethodsHMGEC were cultured in the absence or presence of serum. Sudan III lipid staining, ultrastructural analysis and lipidomic analyses were performed. Impedance sensing, desmoplakin 1/2 mRNA and cytokeratin (CK) 1, 5, 6, 14 levels were evaluated. Serum containing medium supplemented with higher serum, glucose, an omega-3 lipid cocktail, eicosapentaenoic acid or sebomed medium were investigated for lipid accumulation and ultrastructural morphology.ResultsLipid droplet accumulation in HMGEC was induced by serum containing media after 1 day, but decreased over time. Cultivation in serum induced desmosome and cytokeratin filament formation. Desmoplakin 1/2 gene levels were significantly upregulated after 1d of serum treatment. Furthermore, the normalized impedance increased significantly. Lipidome analysis revealed high levels of phospholipids (over 50%), but very low levels of wax ester and cholesteryl esters (under 1%). Stimulation with eicosapentaenoic acid increased lipid accumulation after one day.ConclusionSerum treatment of HMGEC caused lipid droplet formation to some extent but also induced keratinization. The cells did not produce typical meibum lipids under these growth conditions. HMGEC are well suited to study (hyper)keratinization processes of meibomian gland epithelial cells in vitro.
Highlights
Lipid droplet accumulation in human meibomian gland epithelial cell line (HMGEC) was induced by serum containing media after 1 day, but decreased over time
Meibomian gland dysfunction came into focus because most patients suffering from dry eye disease have abnormal meibomian gland function [1]
Materials and Methods Culture of human meibomian gland epithelial cells Experiments were conducted using the human meibomian gland epithelial cell line (HMGEC) that we received as a kind gift by David Sullivan (Schepens Eye Research Institute) and was first described in 2010 [2]
Summary
Meibomian gland dysfunction came into focus because most patients suffering from dry eye disease have abnormal meibomian gland function [1]. The meibomian gland is a holocrine sebaceous gland that produces meibum, an oily secretion that forms the outer layer of the tear film. The meibomian gland consists of basal proliferating cells that show no lipid accumulation whereas during ascent through different stages of maturation cells accumulate lipids progressively until the cell ruptures. After disintegration of mature meibocytes, the released cytoplasm, cell membranes and accumulated lipids form the meibum. The recent development of a meibomian gland epithelial cell line (HMGEC) enables the physiology and pathophysiology of meibocytes to be studied in vitro [2]. The method for handling and cultivation of these immortalized cells is not fully elucidated. To use the meibomian gland epithelial cell line as a model to investigate effects on physiological maturation of meibocytes, maturation in culture must be characterized further
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