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Abstract
Complete loss of the corneal-limbal epithelium leads to re-epithelialisation by bulbar
conjunctival cells. Since conjunctival and corneal-limbal epithelial cells represent
two different cell lines, this conjunctival healing of the cornea is followed by stromal
scarring, decreased visual acuity, and severe discomfort. Unilateral corneal-limbal
epithelial defects can be resolved by the transplantation of limbal grafts taken from
the uninjured eye. However, this procedure requires a large limbal graft to be taken
from the healthy eye, and is not possible for bilateral lesions. We investigated the
possibility of restoring the human corneal surface with autologous corneal epithelial
sheets generated by serial cultivation of limbal cells.
Cells were cultivated from a 1 mm2 biopsy sample taken from the limbus of the healthy
eye of two patients with severe alkali burns, and thus complete loss of the corneal-limbal
surface, of one eye. Normal corneal differentiation was tested with a specific biochemical
marker. Autologous cultured corneal sheets were then grafted onto the damaged eyes
of the two patients. The patients were followed up at more than 2 years after grafting.
We have shown that corneal progenitor cells are localised in the limbus, that cultured
limbal cells generate cohesive sheets of authentic corneal epithelium, and that autologous
cultured corneal epithelium restored the corneal surface of two patients with complete
loss of the corneal-limbus epithelium. Long-term follow-up showed the stability of
regenerated corneal epithelium and the striking improvement in patients' comfort and
visual acuity.
The cultivation of corneal epithelium might offer an alternative to patients with
unilateral lesions and a therapeutic chance to patients with severe bilateral corneal-limbal
epithelial defects. Our findings give a new perspective on the treatment of ocular
disorders characterised by stem-cell deficiency.