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Objective:To construct the tissue engineering seed cell(HaCaT cell line) with stable expression of the human epidermal growth factor(EGF), and analyze the changes of its biological characteristics.Methods:PCDNA3.1-EGF eukaryotic expression vector was transferred intoHaCaT cell, andG418 was utilized to select theHaCaT-EGF cell line.Using an inverted microscope,PCR,ELISA method to detect the changes of the cell morphology, the expression of theEGF gene and protein, and the mRNA expression levels of apoptosis related molecule Caspase-3, the cell cycle related protein cyclinD1.Results:The mRNA expression levels of the obtainedHaCaT-EGF cell were more than100 times higher than the level of ordinaryHaCaT cell.The colony of theHaCaT-EGF cells was more focused and tight compared to the empty vector transfectedHaCaT cells and normalHaCaT cells.The expression levels of apoptotic factor Caspase-3 and cyclinD1 inHaCaT-EGF cell were significantly higher than those in the empty vectorHaCaT- pcDNA3.1 cell, and the differences were statistically significant(P<0.01), but there was no significant difference compared to the normalHaCaT cells(P>0.05).Conclusions:HaCaT-EGF cell can continuously secreteEGF, and the biological characteristic is stable.It can be used for tissue engineering experiment and is an ideal seed cell for constructing tissue engineered skin.