Human skin equivalents expressing exogenous polypeptides

Abstract

The present invention relates generally to compositions for wound closure. More specifically, the present invention provides human skin equivalents engineered to express exogenous polypeptides (e.g., antimicrobial polypeptides and keratinocyte growth factor 2) and compositions and methods for making human skin equivalents engineered to express exogenous polypeptides. In addition, the present invention provides methods for treatment of wounds with human skin equivalents engineered to express exogenous polypeptides.

What is claimed is: 
     
         1 . An in vitro human skin equivalent comprising a stratified squamous epithelium containing immortalized keratinocytes, the immortalized keratinocytes stably transfected with a vector comprising a keratinocyte specific promoter operably linked to a DNA sequence encoding keratinocyte growth factor-2 (KGF-2), wherein said in vitro human skin equivalent is characterized in having correct tissue architecture including the formation of desmosomes, hemidesmosomes, and basal lamina. 
     
     
         2 . The in vitro human skin equivalent of  claim 1 , wherein said keratinocyte specific promoter is an involucrin promoter or a K14 promoter. 
     
     
         3 . The in vitro human skin equivalent of  claim 1 , wherein said immortalized keratinocytes are selected from the group consisting of near-diploid immortalized keratinocytes (NIKS) cells and cells derived from NIKS cells. 
     
     
         4 . The in vitro human skin equivalent of  claim 1 , wherein said expression vector further comprises a selectable marker. 
     
     
         5 . The in vitro human skin equivalent of  claim 1 , wherein said KGF-2 is full-length KGF-2. 
     
     
         6 . A method of treating a subject with a wound, the method comprising contacting the wound of the subject with an in vitro human skin equivalent of  claim 1 . 
     
     
         7 . The method of  claim 6 , wherein said contacting comprises topical application, engraftment or wound dressing. 
     
     
         8 . The method of  claim 6 , wherein said wounds are selected from venous ulcers, diabetic ulcers, pressure ulcers, burns, ulcerative colitis, mucosal injuries, internal injuries, or external injuries. 
     
     
         9 . The method of  claim 6 , wherein said keratinocytes of the in vitro human skin equivalent are mixed with cells obtained from said subject or derived therefrom prior to contacting the wound of the subject with an in vitro human skin equivalent.