Establishing and characterizing a limbal stem cell deficiency mouse model

Session Details

Session Title: Cornea Medical I

Session Date/Time: Monday 12/09/2016 | 17:00-18:30

Paper Time: 17:00

Venue: Hall C2

First Author: : L.Espandar USA

Co Author(s): :    J. Vasseur                    

Abstract Details

Purpose:

To establish a robust mouse model for limbal stem cell deficiency.

Setting:

University of Pittsburgh, USA

Methods:

Right eye of 20 female C57Bl/6 mice underwent 100% ethanol exposure for 30 seconds, copious irrigation then total corneal debridement by Alger brush. 5 Mice were euthanized at each time point of 2h, 3d, 2w, and 4w after injury. Mice underwent clinical evaluation of corneal opacity, epithelial defect and quantification of corneal neovascularization. Nanostring was used for RNA expression profile and confocal microscopy to show changes in staining pattern of different cytokeratin proteins. Histopathology was used to show changes in corneal structure and inflammatory cell recruitment.

Results:

Limbal stem cell deficiency was produced in all eyes. Epithelial defect was smaller in 2-3 days but persistent during 4 weeks follow-up, corneal neovascularization appeared after 3 days. Confocal microscopy showed the presence of k14, k8 progenitor cells in the limbus after injury and increase in their numbers, in 3 days. Cluster of k14/8 positive cells can be appreciated in central cornea after 1 week of injury. Histopathology showed disorganized collagen and thickening of corneal stroma after 1 week of injury, infiltration of neutrophils at 2 days and mononuclears after 1 week. Nanostring data showed up regulation of inflammatory mediators, progenitor cells and Wnt pathway

Conclusions:

We successfully establish and characterized a mouse model of limbal stem cell deficiency. This model can be used to study therapeutic effect of different modalities such as drugs or stem cells in limbal stem cell deficiency.

Financial Disclosure:

NONE