Methods for characterization/manipulation of human corneal stem cells and their applications in regenerative medicine.

Cell therapy is an emerging therapeutic strategy aimed at replacing or repairing severely damaged tissues
with cultured cells. Speci fi cally, ocular burns cause depletion of limbal stem cells, which leads to corneal
opaci fi cation and visual loss. Corneal stem cells are segregated in the basal layer of the limbus, which is the
transitional zone of the epithelium located between the cornea and the bulbar conjunctiva. Autologous
cultured limbal epithelial cells can restore damaged corneas. We sought to establish a culture system that
allows preservation of limbal stem cells and preparation of manageable epithelial sheets. We outline some
quality criteria, which assure the clinical performance of keratinocyte culture: evaluation of the number of
holoclones within a cultured epithelial graft, proportion of aborting colonies, and percentage of cells
expressing high levels of D Np63 a .