June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Evaluation of a protocol for isolation and expansion of murine and rat lacrimal gland mesenchymal stem cells
Author Affiliations & Notes
  • Mathias Roth
    Ophthalmology, University of Düsseldorf, Düsseldorf, Germany
  • Alexander Kunze
    Ophthalmology, University of Düsseldorf, Düsseldorf, Germany
  • Gerd Geerling
    Ophthalmology, University of Düsseldorf, Düsseldorf, Germany
  • Stefan Schrader
    Ophthalmology, University of Düsseldorf, Düsseldorf, Germany
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 5424. doi:
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      Mathias Roth, Alexander Kunze, Gerd Geerling, Stefan Schrader; Evaluation of a protocol for isolation and expansion of murine and rat lacrimal gland mesenchymal stem cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):5424.

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

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Abstract

Purpose: Dry Eye disease is caused by inadequate quantity or quality of tears. One of the major reasons for severe Dry Eye is aqueous tear deficiency due to lacrimal gland insufficiency. Tools to facilitate regeneration of aggrieved lacrimal glands would be a desirable treatment option for patients suffering from severe Dry Eye. Aim of the present study was to establish a standardized protocol for isolation and expansion of putative mesenchymal stem cells (MSC’s) from native murine and rat lacrimal glands to characterize the cells and to assess the differentiation potential in vitro.

Methods: Lacrimal glands were removed from wildtype rats and mice. One part of the tissue was used for cell culture, the other part was embedded for immunostaining. Cultivated cells and lacrimal gland tissue was evaluated for the expression of mesenchymal progenitor cell markers, such as, nestin, thy 1, PDGFR-β, CD146, SMA, desmin, vimentin, GFAP, musashi-1, CD34 and pan-cytokeratin.

Results: Cells expanded from lacrimal gland tissue explants showed a fibroblast-like cell shape and noticeable intracellular lipid droplets. After eight days of culture a subpopulation of cells showed signs of differentiation into myofibroblasts by demonstrating typical stress fibers. Expression of mesenchymal stem cell markers thy-1, PDGFR-β, SMA, desmin and vimentin, musashi-1 and CD146 was demonstrated by immunostaining. Cells appeared to be negative for CD34. Furthermore cells positive for GFAP and nestin could be detected in lacrimal gland tissue sections.

Conclusions: Murine and rat lacrimal gland cells can be successfully isolated and expanded in vitro using explant culture techniques. The outgrowing cell population shows characteristics of mesenchymal stem cells, however more studies are needed to evaluate the differentiation potential and regenerative ability of this cell population.

Keywords: 576 lacrimal gland • 721 stem cells • 687 regeneration  
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