July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Melanocytes as an emerging key player in niche regulation of limbal stem cells
Author Affiliations & Notes
  • Ursula Schlotzer-Schrehardt
    Department of Ophthalmology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Naresh Polisetti
    Department of Ophthalmology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Matthias Zenkel
    Department of Ophthalmology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Elisabeth Naschberger
    Department of Surgery, Division of Molecular and Experimental Surgery, University of Erlangen-Nuernberg, Erlangen, Germany
  • Lukas Heger
    Department of Dermatology, Laboratory of Dendritic Cell Biology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Diana Dudziak
    Department of Dermatology, Laboratory of Dendritic Cell Biology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Michael Stuerzl
    Department of Surgery, Division of Molecular and Experimental Surgery, University of Erlangen-Nuernberg, Erlangen, Germany
  • Friedrich E Kruse
    Department of Ophthalmology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Footnotes
    Commercial Relationships   Ursula Schlotzer-Schrehardt, None; Naresh Polisetti, None; Matthias Zenkel, None; Elisabeth Naschberger, None; Lukas Heger, None; Diana Dudziak, None; Michael Stuerzl, None; Friedrich Kruse, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 3453. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Ursula Schlotzer-Schrehardt, Naresh Polisetti, Matthias Zenkel, Elisabeth Naschberger, Lukas Heger, Diana Dudziak, Michael Stuerzl, Friedrich E Kruse; Melanocytes as an emerging key player in niche regulation of limbal stem cells. Invest. Ophthalmol. Vis. Sci. 2018;59(9):3453. doi: https://doi.org/.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Limbal epithelial stem/progenitor cells (LEPC) and melanocytes have been shown to form structural units in the human limbal stem cell niche (Polisetti et al. 2016). Here, we explored the specific role of melanocytes for LEPC function and niche homeostasis.

Methods : Primary cultures of limbal melanocytes were established and characterized by flow cytometry and immunocytochemistry. Melanocytes were used as feeder layers in direct and indirect LEPC co-culture experiments using 3T3 fibroblasts and limbal mesenchymal stromal cells (MSC) as controls. Co-cultures were analyzed by light and electron microscopy, immunolabeling, qPCR and Western blotting. Colony forming efficiency, cell migration, proliferation, and monocyte adhesion were analyzed by appropriate assays. Limbal epithelial cell sheets were generated on fibrin-based hydrogels with or without melanocytes. The effect of melanocytes on proliferation of T cells and human umbilical vein endothelial cells was investigated using mixed lymphocyte reaction (MLR) and proliferation/tube formation assays.

Results : LEPC co-cultivated with melanocytes showed increased colony growth compared to co-cultivation with 3T3 fibroblasts and MSC. Melanocytes infiltrated the epithelial clones to closely enwrap LEPC with their cell processes and to transfer melanosome-rich packages via phagocytosis. Melanocytes also suppressed mRNA and protein expression of differentiation markers (K3, K12) and upregulated expression of progenitor markers (K15, N-Cadherin, ABCG2) in LEPC, both in direct and indirect co-cultures. Melanocytes also supported LEPC migration and proliferation in 2D- and 3D-organ culture models, and supported epithelial sheet formation on fibrin gels. Stimulation with IFN-γ significantly increased the adhesion of monocytic cells to melanocytes via upregulation of ICAM-1. Most importantly, melanocytes markedly suppressed proliferation of activated T cells and vascular endothelial cells through direct and indirect effects, which exceeded those of MSC.

Conclusions : These findings support the concept that melanocytes, apart from melanogenesis and melanosome transfer, control LEPC homeostasis in the limbal niche and contribute to the integrity of the limbal barrier. Based on their effective immunomodulatory and anti-angiogenic properties, limbal melanocytes have great potential for LEPC ex vivo-expansion and therapeutic application.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

×
×

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.

×