Molecular Therapy
Volume 10, Issue 1, July 2004, Pages 86-96
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Article
Accelerated Wound Healing by In vivo Application of Keratinocytes Overexpressing KGF

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Abstract

Epidermal regeneration is a complex process, strongly influenced by growth factors, including keratinocyte growth factor (KGF). The objective of this study was to establish immortalized HaCaT keratinocytes and KMST-6-fibroblasts stably expressing KGF. Transfection efficiency, genomic integration, and functionality of the transgene were determined by ELISA and PCR, and KGF-expressing clones were selected using an air–liquid interface test system. HaCaT cells displayed stronger transgene expression compared to transfected fibroblasts, and the most effective HaCaT clone was incubated on a membrane carrier to form a “membrane cell graft.” Twenty-one superficial second-degree burn wounds were created in each of three pigs, and wound healing capacity of the generated “polypeptide cell delivery system” after grafting was examined. Untransfected HaCaT keratinocytes and membrane-covered and untreated burn wounds served as controls. Histological and macroscopical follow-up revealed that grafting of transfected HaCaT cells resulted in complete reepithelialization within 5 days, while wounds covered with untransfected cells needed 2 days longer. At untreated sites, a thin epithelium was detectable after 10 days. The results indicate that wound healing processes can be stimulated distinctly by growth factors secreted from HaCaT cells, with a prominent role for transgenic KGF.

Keywords

growth factors
wound healing
gene transfer techniques
molecular therapy
transformed cell line
transplantation

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These authors contributed equally to this work.