Objective To provide a new way for treatment of full thickness skin defects by embryonic stem (ES) cellderived epidermallike stem cells. Methods Epidermal like stem cells, labeled by Hoechst 33342 and carried by a layer of biomembrane, were transplanted into the defective skin of mice. The differentiation tissue of donor cells was sampled each week. The sections were observed with HE staining, immunohistochemical and dilabeled immunofluorescence methods to test the expression of β1 integrin, CK15, CK19, CK10, CEA. Results The full thickness skin defects were healed in 2 weeks. The newborn skin was thicker than the normal skin. The basal layer cells proliferated.There were more bulky cellular poles towards dermis. The cells labeled by Hoechst 33342, located in the newborn epidermis and tubular or follicular structures in dermis, expressed β1 integrin and CK15 positive respectively in the first 3 weeks. There were sweat glandlike, sebaceous glandlike and hair folliclelike structures in the newborn dermis after 4 weeks. Basal cells of keratinized stratified squamous epithelium expressed CK19 and CK10 positive respectively and sweat glandlike structure expressed CEA positive. Conclusion ES cellderived epidermal stem cell