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Last Updated: March 29, 2024

Claims for Patent: 8,871,505


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Summary for Patent: 8,871,505
Title:Artificial skin
Abstract: The present invention relates to a method for producing artificial skin, comprising: adding a matrix metalloproteinase inhibitor and a heparanase inhibitor to an artificial skin formation culture medium comprising human epidermal keratinocytes and human dermal fibroblasts, culturing the cells in the artificial skin formation culture medium, and forming artificial skin.
Inventor(s): Iriyama; Shunsuke (Yokohama, JP), Umishio; Kenichi (Yokohama, JP), Tsunenaga; Makoto (Yokohama, JP), Inomata; Shinji (Yokohama, JP), Adachi; Eijiro (Yokohama, JP)
Assignee: Shiseido Company, Ltd. (Tokyo, JP)
Application Number:13/582,199
Patent Claims:1. A method for producing artificial skin, comprising: adding a matrix metalloproteinase inhibitor and a heparanase inhibitor to an artificial skin formation culture medium comprising human epidermal keratinocytes and human dermal fibroblasts, culturing the cells in the artificial skin formation culture medium, and forming artificial skin, wherein the artificial skin comprises an epidermal basement membrane containing a continuous lamina densa and anchoring fibers arising from the lamina densa, and a dermis containing collagen fibers, and wherein the anchoring fibers arising from the lamina densa are securely bonded to the collagen fibers in the dermis.

2. The method according to claim 1, wherein the matrix metalloproteinase inhibitor is an inhibitor of a matrix metalloproteinase selected from the group consisting of gelatinase, collagenase, stromelysin, and matrilysin.

3. The method according to claim 1, wherein the matrix metalloproteinase inhibitor is selected from the group consisting of N-hydroxy-2-[[(4-methoxyphenyl)sulfonyl](3-picolyl)amino]-3-methylbutanam- ide hydrochloride and p-NH.sub.2-Bz-Gly-Pro-D-Leu-Ala-NHOH.

4. The method according to claim 1, wherein the matrix metalloproteinase inhibitor is added to the culture medium in an amount from about 10 .mu.g/L to 10 g/L.

5. The method according to claim 4, wherein the matrix metalloproteinase inhibitor is added to the culture medium in an amount from about 100 .mu.g/L to 1 g/L.

6. The method according to claim 5, wherein the matrix metalloproteinase inhibitor is added to the culture medium in an amount from about 1 mg/L to 100 mg/L.

7. The method according to claim 1, wherein the heparanase inhibitor is 1-[4-(1H-benzoimidazol-2-yl)-phenyl]-3-[4-(1H-benzoimidazol-2-yl)-phenyl]- -urea.

8. The method according to claim 1, wherein the heparanase inhibitor is added to the culture medium in an amount from about 10 .mu.g/L to 100 g/L.

9. The method according to claim 8, wherein the heparanase inhibitor is added to the culture medium in an amount from about 100 .mu.g/L to 10 g/L.

10. The method according to claim 9, wherein the heparanase inhibitor is added to the culture medium in an amount from about 1 mg/L to 1 g/L.

11. The method according to claim 1, wherein the culture medium is selected from the group consisting of (i) Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serume; (ii) DMEM-Ham's F-12 (3:1) medium containing 10% fetal bovine serum, 5 .mu.g/ml transferrin, 5 .mu.g/ml insulin, 2 nM triiodothyronine, 0.1 nM cholera toxin, and 0.4 .mu.g/ml hydrocortisone; and (iii) a 1:1 mixture of keratinocyte growth medium and DMEM containing 10% fetal bovine serum.

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