Claims
- 1. A method for examining the effect of a test factor on human skin equivalent, comprising the steps of:
providing a human stratified squamous epithelial cell culture, wherein the culture comprises spontaneously immortalized human keratinocytes and forms a reconstructed epidermis; exposing the reconstructed epidermis to the factor; and evaluating the effect of the factor on the reconstructed epidermis.
- 2. The method of claim 1, wherein the spontaneously immortalized human keratinocytes are ATCC CRL-12191 cells.
- 3. The method of claim 1, wherein the spontaneously immortalized human keratinocytes carry an exogenous gene.
- 4. The method of claim 3, wherein the transgene is green fluorescence protein gene.
- 5. The method of claim 1, wherein the factor is a material that may contact human skin.
- 6. The method of claim 5, wherein the material is selected from the group consisting of compounds and mixture of compounds.
- 7. The method of claim 6, wherein the compound is selected from the group consisting of therapeutics, cosmeceuticals or cosmetic skin products.
- 8. The method of claim 6, wherein the mixture of compounds is selected from the group consisting of therapeutics, cosmeceutical products or cosmetic skin products.
- 9. The method of claim 5, wherein the material is selected from the group consisting of inorganic acids, organic acids, acid mixtures, acid derivatives, amines, inorganic bases, organic bases, base mixtures, a mixture of an acid and a base, cleaners, detergents, surfactants, and industrial chemicals.
- 10. The method of claim 1, wherein the factor is selected from the group consisting of UV light, radiation, air pressure, environment temperature and friction.
- 11. The method of claim 1, wherein the effect is selected from the group consisting of cell differentiation, cell proliferation, cell survival, cell damage and cell death.
- 12. The method of claim 1, wherein the effect is selected from the group consisting of tissue culture morphology change, barrier function and tissue strength.
- 13. The method of claim 1, wherein the effect is toxicity.
- 14. The method of claim 1, wherein the culture further comprises a base layer of collagen and fibroblasts.
- 15. The method of claim 1, wherein the factor is in liquid form and is exposed to the reconstructed epidermis as a liquid.
- 16. The method of claim 1, wherein the factor is in solid form and is exposed to the reconstructive epidermis as a solid.
- 17. The method of claim 1, wherein the factor is in emulsion form and is exposed to the epidermis as an emulsion.
- 18. The method of claim 1, wherein the factor is combined with a solvent and exposed to the reconstructed epidermis as a solvent/factor combination.
- 19. The method of claim 1, wherein the agent being screened is in liquid form and is exposed to the reconstructed epidermis as a liquid.
- 20. The method of claim 1, wherein the agent being screened is in solid form and is exposed to the reconstructed epidermis of the solid.
- 21. The method of claim 1, wherein the agent being screened is an emulsion and is exposed to the reconstructed epidermis as an emulsion.
- 22. The method of claim 1, wherein the agent being screened is combined with the solvent and is exposed to the reconstructed epidermis as a solvent/agent combination.
- 23. The method of claim 1, wherein the evaluating comprises microscopic analysis of the reconstructed epidermis.
- 24. The method of claim 1, wherein the evaluating comprises a histological examination of the reconstructed epidermis.
- 25. The method of claim 1, wherein the evaluating comprises a visual examination of the reconstructed epidermis.
- 26. The method of claim 1, wherein the evaluating comprises cell viability analysis.
- 27. A method for selecting preventive or therapeutic agents for skin damage or benefits caused by a factor, comprising the steps of:
providing a human stratified squamous epithelial cell culture, wherein the culture comprises spontaneously immortalized human keratinocytes and forms a reconstructed epidermis; exposing the reconstructed epidermis to a factor; exposing the reconstructed epidermis to an agent being screened for preventive or therapeutic use; and evaluating the effect of the agent on the reconstructed epidermis.
- 28. The method of claim 27, wherein exposing the reconstructed epidermis to the factor occurs prior to exposing the reconstructed epidermis to an agent being screened for preventive or therapeutic use.
- 29. The method of claim 27, wherein exposing the reconstructed epidermis to the factor occurs after exposing the reconstructed epidermis to an agent being screened for preventive or therapeutic use.
- 30. The method of claim 27, wherein exposing the reconstructed epidermis to the factor and exposing the reconstructed epidermis to an agent being screened for preventive or therapeutic use occur at the same time.
- 31. The method of claim 27, wherein the factor is in liquid form and is exposed to the reconstructed epidermis as a liquid.
- 32. The method of claim 27, wherein the factor is in solid form and is exposed to the reconstructive epidermis as a solid.
- 33. The method of claim 27, wherein the factor is in emulsion form and is exposed to the epidermis as an emulsion.
- 34. The method of claim 27, wherein the factor is combined with a solvent and exposed to the reconstructed epidermis as a solvent/factor combination.
- 35. The method of claim 27, wherein the agent being screened is in liquid form and is exposed to the reconstructed epidermis as a liquid.
- 36. The method of claim 27, wherein the agent being screened is in solid form and is exposed to the reconstructed epidermis of the solid.
- 37. The method of claim 27, wherein the agent being screened is an emulsion and is exposed to the reconstructed epidermis as an emulsion.
- 38. The method of claim 27, wherein the agent being screened is combined with the solvent and is exposed to the reconstructed epidermis as a solvent/agent combination.
- 39. The method of claim 27, wherein the evaluating comprises microscopic analysis of the reconstructed epidermis.
- 40. The method of claim 27, wherein the evaluating comprises a histological examination of the reconstructed epidermis.
- 41. The method of claim 27, wherein the evaluating comprises a visual examination of the reconstructed epidermis.
- 42. The method of claim 27, wherein the evaluating comprises cell viability analysis.
- 43. A method of evaluating the effects of a plurality of compounds on a human skin equivalent comprising:
(a) providing a plurality of cell culture comprising spontaneously immortalized human keratinocytes and a set of test compounds; (b) sequentially exposing said plurality of cell cultures to said set of compounds alone or in combination; and (c) evaluating the effects of said set of compounds on said cell cultures.
- 44. The method of claim 43, wherein said cell cultures are provided in formal selected from the group consisting of 24 well plates and 96 well plates.
- 45. The method of claim 43, wherein said set of compounds is combinatorial library of compounds.
- 46. The method of claim 43, wherein said spontaneously immortalized human keratinocytes comprise a reporter gene.
- 47. The method of claim 46, wherein said reporter gene is GFP.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part application of U.S. Ser. No. 09/769,124, filed on Jan. 24, 2001, which is a continuation of U.S. Ser. No. 09/277,295, filed on Mar. 26, 1999, which is a continuation of U.S. Ser. No. 09/114,557, filed on Jul. 13, 1998 (U.S. Pat. No. 5,989,837, Nov. 23, 1999). These applications are incorporated by reference herein.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
[0002] This invention was made with United States government support awarded by the following agencies: NIH Grant No(s). AR40284, CA66786 and AR42853. The United States has certain rights in this invention.
Continuations (2)
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Number |
Date |
Country |
Parent |
09277295 |
Mar 1999 |
US |
Child |
09769124 |
Jan 2001 |
US |
Parent |
09114557 |
Jul 1998 |
US |
Child |
09277295 |
Mar 1999 |
US |
Continuation in Parts (1)
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Number |
Date |
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Parent |
09769124 |
Jan 2001 |
US |
Child |
09945136 |
Aug 2001 |
US |