Objective   To explore the differences of human epidermal stem cells (ESCs) and human immortalized keratinocytes HaCaT as seed cells to construct tissue engineered skin.    Methods   Human ESCs were selected by rapid attachment to collagen Ⅳ

and fed with DK-SFM. The cells at passages 2 were ready to seed. Hacat cells were also fed with DK-SFM. ESCs and Hacat cells were respectively seeded on collagen matrices populated with increasing numbers of fibroblasts and cultured for 1 week at the air liquid interface. Morphology

physical properties observation

and HE staining were carried out to compare the differences of the 2 cells.    Results   ESCs were faster in constructing a contraction of the tissue-engineered skin

and had good tension and toughness and similar stratification as normal skin. HaCaT cells were less laminated

slow growth

and had low tension and toughness in tissue-engineered skin construction. The diameter of 2 types of constructed skin samples was shortened along with the increasing of culture time

and had significant difference in 3 to 7 d.    Conclusion   Epidermal stem cells are more suitable as seed cells for tissue engineering of skin than the HaCaT immortalized cells.