Human skin equivalents expressing exogenous polypeptides

1. A composition comprising a human skin equivalent comprising stratified Near-diploid Immortalized Keratinocyte (NIKS) cells adhered to a physiological substratum of dermal fibroblasts embedded within a fibrillar collagen base, said NIKS cells expressing a heterologous TIMP-1 gene and having normal cellular proliferation as compared to untransfected Near-diploid Immortalized Keratinocyte cells, wherein said gene encoding said heterologous TIMP-1 gene is operably linked to a promoter sequence that allows TIMP-1 gene expression in said human skin equivalent, wherein said promoter sequence is selected from the group consisting of a K14 promoter, an involucrin promoter, and a ubiquitin promoter.

2. A method for providing cells expressing a heterologous TIMP-1, comprising:

a) providing Near-diploid Immortalized Keratinocyte (NIKS) cells and an expression vector comprising a DNA sequence encoding a TIMP-1 gene operably linked to a regulatory sequence;

b) introducing said expression vector to said Near-diploid Immortalized Keratinocyte cells; and

c) organotypically culturing said Near-diploid Immortalized Keratinocyte cells under conditions such that said cells stratify and adhere to a physiological substratum of dermal fibroblasts embedded within a fibrillar collagen base and said heterologous TIMP-1 is expressed and wherein said Near-diploid Immortalized Keratinocyte cells have normal cellular proliferation as compared to untransfected Near-diploid Immortalized Keratinocyte cells.

3. The method of claim 2 , wherein said Near-diploid Immortalized Keratinocyte cells are capable of stratifying into squamous epithelia.

4. The method of claim 2 , further comprising co-culturing said Near-diploid Immortalized Keratinocyte cells with cells derived from a patient.

5. A method of treating skin wounds comprising:

a) providing a human skin equivalent comprising stratified Near-diploid Immortalized Keratinocyte cells (NIKS) cells adhered to a physiological substratum of dermal fibroblasts embedded within a fibrillar collagen base, said NIKS cells expressing a heterologous TIMP-1 gene wherein said Near-diploid Immortalized Keratinocyte cells have normal cellular proliferation as compared to untransfected Near-diploid Immortalized Keratinocyte cells, and a subject with a wound;

b) contacting said skin wound with said human skin equivalent expressing a heterologous TIMP-1 gene under conditions such that said wound is treated.

6. The method of claim 5 , wherein said contacting comprises a technique selected from the group consisting of topical application, engraftment and application of a wound dressing.

7. The method of claim 5 , wherein said skin wounds are selected from the group comprising venous ulcers, diabetic ulcers, pressure ulcers, burns, and external injuries.