April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Expansion of Limbal Progenitors on Type IV Collagen
Author Affiliations & Notes
  • N. Szentmary
    Dept of Ophthalmology, Semmelweis University, Budapest, Hungary
  • K. Barabás
    Laboratory of Cellular and Developmental Neurobiology, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary
  • A. Stündl
    Dept of Ophthalmology, Semmelweis University, Budapest, Hungary
  • K. Demeter
    Laboratory of Cellular and Developmental Neurobiology, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary
  • J. Németh
    Dept of Ophthalmology, Semmelweis University, Budapest, Hungary
  • I. Süveges
    Dept of Ophthalmology, Semmelweis University, Budapest, Hungary
  • E. Madarász
    Laboratory of Cellular and Developmental Neurobiology, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary
  • Footnotes
    Commercial Relationships  N. Szentmary, None; K. Barabás, None; A. Stündl, None; K. Demeter, None; J. Németh, None; I. Süveges, None; E. Madarász, None.
  • Footnotes
    Support  OTKA Grant T048477
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 1785. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      N. Szentmary, K. Barabás, A. Stündl, K. Demeter, J. Németh, I. Süveges, E. Madarász; Expansion of Limbal Progenitors on Type IV Collagen. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1785.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : To determine immunocythochemical properties and proliferative potential of limbal stem-like cells maintained in vitro, on collagen.

Methods: : Limbal pieces were harvested from multiorgan donors and cultured on collagen IV using culture media conditioned by embryonic mouse fibroblasts (E16). Immunocytochemical properties were determined by p63, cytokeratin 19 (CK19), ssea-3 and bcrp (ABCG-2) stainings, cells with active metabolism by MTT assay (3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromid -> formazan) (0.25mg/ml) and the percentage of proliferating cells by propidium iodid uptake (FACS).

Results: : 24-48 hours following explantation onto type IV collagen, spheroid cells grew out of the explants. After 48 hours, migrating bipolar and epitheloid cells appeared. After 3-4 days, the center of the explants was necrotized. 80-90% of the cells in the outgrowth zone were viable and about 7 % of non-attached cells was in S or G2 phase. A number of outgrowing cells showed p63, CK19, ssea-3 and bcrp immune-reactivity.

Conclusions: : Cells growing out from limbal explants on collagen, show immunocytochemical characteristics of limbal stem cells. Collagen-like surfaces may be the appropriate substrates for ex vivo expansion and for a future in vivo implantation of limbal stem cells.

Keywords: cornea: basic science • cornea: epithelium • cornea: clinical science 
×
×

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.

×