April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Characterization of Adult Mouse Lacrimal Gland Explant Cultures
Author Affiliations & Notes
  • Daniela Marcano
    Baylor College of Medicine, Houston, TX
  • Ghanashyam Acharya
    Baylor College of Medicine, Houston, TX
  • Stephen C Pflugfelder
    Baylor College of Medicine, Houston, TX
  • Footnotes
    Commercial Relationships Daniela Marcano, None; Ghanashyam Acharya, None; Stephen Pflugfelder, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 60. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Daniela Marcano, Ghanashyam Acharya, Stephen C Pflugfelder; Characterization of Adult Mouse Lacrimal Gland Explant Cultures. Invest. Ophthalmol. Vis. Sci. 2014;55(13):60.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: Dry eye syndrome is caused by the dysfunction of the lacrimal gland. One of the proposed strategies to treat this pathology is based on organ transplant from an ex-vivo regenerated organ. Decellularization, germ method, culture on 3D Magtrigel systems and culture on nano fibers are techniques that have been used to mimic the cellular matrix in order to regenerate the organ by promoting cell proliferation and differentiation. However, most of these procedures use embryonic or purified acinar cells, which might be difficult to reproduce in humans. The purpose of the study was to establish lacrimal gland explant cultures from adult mice and to characterize the cellular outgrowth. This information will be beneficial for organ regeneration.

Methods: Lacrimal glands from 6-8 week old C57BL/6 mice were aseptically removed, washed in Ham’s media, minced, and digested in liberase. Tissue pieces were plated on plastic dishes and cultured for 8 to 14 days with constant monitoring of morphology and cell differentiation. Subsequently, cultures were washed, fixed with cold methanol, and analyzed by immunofluorescent (IF) microscopy. For comparison, lacrimal gland paraffin sections were also stained by IF. Cytokeratins 5, 7, 8, 14, and 18, vimetin and AQP5 and AQP11 were used as markers. Also, the secretory proteins epidermal growth factor (EGF), lactoferrin, and MUC5AC were evaluated.

Results: Phase contrast images of the cellular outgrowth showed a mixture of elongated cells and polygonal cells. Some binucleated spindle-shape and large flattened cells were also detected. IF showed the elongated cells were vimetin positive. Moreover, the polygonal cells were positive for K5, K8, and K14. Interestingly, lacrimal cultures were repeatedly negative (at least three times) for K7 or K18. However; positive immunstaining for these markers was observed in lacrimal gland paraffin sections. Both explant cultures and paraffin sections were positive for AQP5, AQP11, EGF, lactoferrin and MUC5AC.

Conclusions: A mixture of epithelial (polygonal cells; keratin positive, producing secretory proteins) and mesenchymal cells (elongated cells; vimetin positive) was obtained from murine lacrimal gland explants cultures.

Keywords: 449 cell survival  
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×