PURPOSE To present results of ocular surface reconstruction with application of cultured corneal epithelium in limbal stem cell insufficiency. MATERIAL AND METHODS 25 patients suffered from limbal insufficiency in one eye after a chemical or thermal burn. Their healthy eyes were the source of limbal epithelium for corneal epithelial culture. Limbal cells from a 2 mm2 biopsy were seeded on amniotic membrane. Cultures were carried out in standard conditions in supplemented DMEM in presence of 3T3 fibroblasts. After superficial keratectomy the amniotic carriers with epithelial cells were transplanted on denuded corneas. The presence of corneal vascularization, epithelial regularity and visual acuity were evaluated. RESULTS Postoperative follow-up ranged from 3 up to 9 months, average 4.9 +/- 1.1 months. 3 months after the surgery 72.0% of the eyes showed stable epithelium with slight corneal haze caused by the amnion. In 60.0% of the eyes there was no recurrent conjunctival neovascularization. 28.0% of the eyes remained cloudy due to stromal revascularization. In 2 eyes total conjunctival pannus developed again. Visual acuity ranged from counting fingers to 0.6. CONCLUSION Transplantation of cultured epithelium is a promising method in treatment of limbal stem cell insufficiency giving satisfactory restoration of corneal epithelium.