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Nagayoshi Asano, Ulrike Hampel, Garreis Fabian, Antje Schröder, Martin Schicht, Sabine Möbius, Friedrich P Paulsen; Morphological characterization of a meibomian gland epithelial cell line. Invest. Ophthalmol. Vis. Sci. 2014;55(13):19.
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© ARVO (1962-2015); The Authors (2016-present)
To characterize a meibomian gland epithelial cell line (kind courtesy of David Sullivan, Boston, SERI, MA) and to investigate morphological changes of cultivated meibocytes after treatment with different media supplements.
Meibocytes were grown two and three dimensionally (in a scaffold and with exposure to air (air-lift)) with keratinocyte medium. Differentiation of two dimensionally cultivated meibocytes was induced by differentiation medium (Dulbecco's Modified Eagle's Medium containing epithelial growth factor and 10% fetal calf serum (FCS)) for 1, 3, 7, or 14 days. Furthermore, differentiation medium was complemented with either 20% FCS, omega 3 fatty acid cocktail, eicosapentaenoic acid or high glucose for 1 day or 7 days. Lipid droplets were visualized with Sudan III staining. Ultrastructural changes over differentiation period were investigated by transmission electron microscopy (TEM). Cytokeratin (CK) expression was analyzed by Western blot.
Histological and TEM analysis of two and three dimensionally cultivated meibocytes indicated that cells resembled basal and differentiating meibocytes that were CK5, -10 and -14 positive but did not develop to mature or hypermature meibocytes. Lipid droplet accumulation in differentiating meibocytes was induced by differentiation media after 1 day, but decreased over time. Meibocytes showed highest lipid droplet accumulation after 1 day of 20% FCS supplementation. Omega 3 fatty acid cocktail and eicosapentaenoic acid increased lipid accumulation after 1 day.
Meibocytes of the meibomian gland epithelial cell line reach a state of differentiating meibocytes after treatment with media supplementation, however induction of meibocyte maturation or hypermature cells is limited.
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