Reconstructed Tissue-engineered Skin with ES Cell-derived Epidermal Stem Cells and Collagen Sponge

Objective Using tissue-engineered skin reconstructed with ES cell-derived epidermal stem cells and collagen sponge to restore the full thickness skin defects.Methods The dermal scaffolds were reconstructed by collagen sponge compounded with mice fetus skin fibroblasts.The scaffolds were transplanted into mice full thickness skin defects.ES cell-derived epidermal stem cells,labeled with Hoechst 33342,carried by a layer of biomembrane,were transplanted into the dermal scaffolds.The differentiation tissue of the tissue-engineered skin was sampled each week.The sections were observed with HE staining,immunohistochemical and di-labeled immunofluorescence methods to test β1 integrin,CK15,CK19,CK10,and CEA.Results The full thickness skin defects,covered with tissue-engineered skin,were healed within 3 weeks.The newborn skin was thicker than normal skin.The basal layer of newborn skin was increased.The labeled cells in the epidermis formed much bulky cellular poles towards dermis and expressed β1 integrin,CK15 and CK19.The labeled tubular or follicular structure expressed β1 integrin and CEA in dermis within 2 weeks.The basal cells of keratinized stratified squamous epithelium expressed CK19 and CK10.There were sebaceous glands-like and hair follicles-like structures in newborn skin after 4 weeks.Conclusion Reconstructed tissue-engineered skin with ES cell-derived epidermal stem cells and collagen sponge can restore the mice full thickness skin defect.There are epidermis,sweat glands-like,sebaceous glands-like and hair follicles-like structures in newborn skin.