Abstract
Besides adequate matrix and cell culture technology, gene therapy may become the third pillar of tissue engineering. Can we use genetically modified human allogenic epidermal keratinocytes as a hEGF secretary tissue for wound healing? Human epidermal keratinocyte can grow in serum-free medium and sub-cultured in vitro. In the present experiment, cDNA encoding the human kidney EGF signal peptide and EGF was inserted in-frame into pBK-CMV plasmid. LipofectAMINE-mediated gene transfer was used to introduce the hEGF gene into subcultured and spontaneously immortalized human keratinocytes. The result shows that transfected human keratinocytes can efficiently secrete biologically active hEGF into the culture medium. The clone of gene transfected keratinocyte can be selected by G418 after gene transfer. The selected clone cells still have normal keratinocyte-like morphological shapes under microscopy and the ability to grow and proliferate. The results indicate that the transfected allogenic human keratinocytes may be used as a general vehicle for the delivery of gene products by means of grafting or mixed grafting with cultured autologous keratinocytes to treat severe burns or “non-healing” wounds.
Published Version
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