June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Limbal mesenchymal stromal cells (L-MSC) display immunosuppressive properties across donor and species boundaries
Author Affiliations & Notes
  • Damien Harkin
    School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD, Australia
    Queensland Eye Institute, Brisbane, QLD, Australia
  • Laura Bray
    School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD, Australia
    Queensland Eye Institute, Brisbane, QLD, Australia
  • Celena Heazlewood
    Mater Medical Research Institute, Brisbane, QLD, Australia
  • Kerry Atkinson
    Mater Medical Research Institute, Brisbane, QLD, Australia
  • Footnotes
    Commercial Relationships Damien Harkin, None; Laura Bray, None; Celena Heazlewood, None; Kerry Atkinson, Osiris Therapeutics Inc (I), Mesoblast Ptl (C)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 1001. doi:
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    • Get Citation

      Damien Harkin, Laura Bray, Celena Heazlewood, Kerry Atkinson; Limbal mesenchymal stromal cells (L-MSC) display immunosuppressive properties across donor and species boundaries. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1001.

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

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Abstract

Purpose: We have evaluated the immunosuppressive properties of L-MSC with the view to using these cells in allogeneic cell therapies for corneal disorders. We hypothesized that L-MSC cultures would suppress T-cell activation, in a similar way to those established from human bone marrow (BM-MSC).

Methods: MSC cultures were established from the limbal stroma of cadaveric donor eye tissue (up to 1 week postmortem) using either conventional serum-supplemented growth medium or a commercial serum-free medium optimized for bone marrow derived MSC (MesenCult-XF system). The MSC phenotype was examined by flow cytometry according to current and emerging markers for human MSC. Immunosuppressive properties were assessed using a mixed lymphocyte reaction (MLR) assay, whereby the white cell fraction from two immunologically incompatible blood donors are cultured together in direct contact with growth arrested MSC. T-cell activation (proliferation) was measured by uptake of tritiated thymidine. Human L-MSC were tested in parallel with human BM-MSC and rabbit L-MSC. Human and rabbit L-MSC were also tested for their ability to stimulate the growth of limbal epithelial (LE) cells in colony formation assays (for both human as well as rabbit LE cells).

Results: L-MSC cultures were >95% negative for CD34, CD45 and HLA-DR and positive for CD73, CD90, CD105 and HLA-ABC. Modest levels (30%) of CD146 expression were observed for L-MSC cultures grown in serum-supplemented growth medium, but not those grown in MesenCult-XF. All MSC cultures derived from both human and rabbit tissue suppressed T-cell activation to varying degrees according to culture technique and species (MesenCult-XF >> serum-fed cultures, rabbit L-MSC >> human L-MSC). All L-MSC stimulated colony formation by LE cells irrespectively of the combination of cell species used.

Conclusions: L-MSC display immunosuppressive qualities, in addition to their established non-immunogenic cell surface marker profile, and stimulate LE cell growth in vitro across species boundaries. These results support the potential use of allogeneic or even xenogeneic L-MSC in the treatment of corneal disorders.

Keywords: 484 cornea: stroma and keratocytes • 482 cornea: epithelium • 555 immunomodulation/immunoregulation  
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