Abstract
Purpose:
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.
Methods:
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.
Results:
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.
Conclusions:
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.
Keywords: 526 eyelid •
583 lipids