| Purpose In this study, the optimal condition for induction of differentiation of epithelial progenitor cells from embryonic stem cells (ES) in vitro and in vivo was determined for use in transplantation to damaged cornea in rabbits.
Methods Mouse ES cell line (ES-GFP) were cultured in flasks coated with type Ⅳcollagen, and were pre-induced with retinoic acid(RA). The optimal condition for pre-induction of epithelial progenitor cells from ES-GFP in vitro and in vivo was determined, and the progenitors were transplanted onto rabbit eyes on which corneal epithelia had been damaged by exposing to n-heptanol.
Results Mouse ES-GFP cells were pre-induced by RA for 2 hours, and completely re-epithelialization of the corneal surface occurred within a month after transplantation. The resultant corneal epithelial cells expressed Green Fluorescence Protein (GFP) of the grafted cells, and no teratomata were observed during the following 2 months period.
Conclusions Epithelia were successfully induced in vitro and in vivo from ES cells and were applicable as grafts for treating corneal epithelial injury. ES cells may become an unlimited donor source of corneal epithelial cells for corneal transplantation and may restore useful vision in patients with a deficiency of corneal limbal epithelial cells. This is an important first trial toward differentiating ES cells in vivo on damaged cornea to reconstruct corneal epithelial cells.
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