Persistent corneal epithelial defect (PED) is a consequence of many ocular surface disorders. The treatment of this disease have faced a lot of difficulties up to now. The transplatation of epithelial cells sheets is the new promised method for PED. The study of tissue-cultured human cord lining epithelial cells transplantation was carried out in 31 eyes of PED (31 patients) from January 2011 to December 2012 in Department of Cornea and External diseases, Vietnam National Institute of Ophthalmology. All of these eyes were sufferd from long time corneal ulcer with average time 10,3 + or - 5,4 weeks, and average time of PED 5,2 + or - 2,8 weeks. Tweenty nine of 31 eyes were healed with the cells transplantation and 25 eyes of them healed within 2 weeks postoperatively. There are normal corneal scars and normal corneal epithelial cell (by impression cytology detection) on transplantation site in all 29 successful cases with 3 - 27 months follow-up. Conclusion: tissue-cultured human cord lining epithelial cells transplantation is a quite effective treatment for persistent corneal epithelial defect.