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Serum-induced keratinization processes in an immortalized human meibomian gland epithelial cell line

Journal Article


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Abstract


  • Purpose

    The 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.

    Methods

    HMGEC 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.

    Results

    Lipid 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.

    Conclusion

    Serum 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.

UOW Authors


  •   Hampel, Ulrike (external author)
  •   Schroeder, Antje (external author)
  •   Mitchell, Todd
  •   Brown, Simon (external author)
  •   Snikeris, Peta (external author)
  •   Garreis, Fabian (external author)
  •   Kunnen, Carolina M. E. (external author)
  •   Willcox, Mark D. P. (external author)
  •   Paulsen, Friedrich (external author)

Publication Date


  • 2015

Citation


  • Hampel, U., Schroeder, A., Mitchell, T., Brown, S., Snikeris, P., Garreis, F., Kunnen, C., Willcox, M. & Paulsen, F. (2015). Serum-induced keratinization processes in an immortalized human meibomian gland epithelial cell line. PLoS One, 10 (6), e0128096-1-e0128096-18.

Scopus Eid


  • 2-s2.0-84935009139

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=3953&context=smhpapers

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/2931

Start Page


  • e0128096-1

End Page


  • e0128096-18

Volume


  • 10

Issue


  • 6

Abstract


  • Purpose

    The 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.

    Methods

    HMGEC 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.

    Results

    Lipid 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.

    Conclusion

    Serum 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.

UOW Authors


  •   Hampel, Ulrike (external author)
  •   Schroeder, Antje (external author)
  •   Mitchell, Todd
  •   Brown, Simon (external author)
  •   Snikeris, Peta (external author)
  •   Garreis, Fabian (external author)
  •   Kunnen, Carolina M. E. (external author)
  •   Willcox, Mark D. P. (external author)
  •   Paulsen, Friedrich (external author)

Publication Date


  • 2015

Citation


  • Hampel, U., Schroeder, A., Mitchell, T., Brown, S., Snikeris, P., Garreis, F., Kunnen, C., Willcox, M. & Paulsen, F. (2015). Serum-induced keratinization processes in an immortalized human meibomian gland epithelial cell line. PLoS One, 10 (6), e0128096-1-e0128096-18.

Scopus Eid


  • 2-s2.0-84935009139

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=3953&context=smhpapers

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/2931

Start Page


  • e0128096-1

End Page


  • e0128096-18

Volume


  • 10

Issue


  • 6