Skin substitutes for irritancy testing

1. A method of screening compounds suspected of being skin irritants, comprising:

a) providing

i) a compound suspected of being a skin irritant;

ii) a skin equivalent comprising a irritant responsive reporter gene construct, wherein said construct comprises a promoter selected from the group consisting of interleukin-8 and interleukin-1α promoters operably linked to a reporter gene; and

b) contacting said skin equivalent with said compound; and

c) measuring the level of gene expression from said reporter gene construct, wherein increased expression from said reporter gene construct is indicative of a response to said irritant.

2. The method of claim 1 , wherein said human skin equivalent has a surface electrical capacitance of from about 80 to about 120 pF.

3. The method of claim 1 , wherein the content of ceramides 5, 6, and 7 in said skin equivalent is from about 20 to about 50% of total ceramide content.

4. The method of claim 1 , wherein the content of ceramide 2 in said skin equivalent is from about 10 to about 40% of total ceramide content.

5. The method of claim 1 , wherein said skin equivalent comprises keratinocytes selected from the group consisting of primary keratinocytes and immortalized keratinocytes.

6. The method of claim 5 , wherein said immortalized keratinocytes are NIKS cells.

7. The method of claim 1 , wherein said keratinocytes express heterologous Kruppel-like factor 4.