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Last Updated: May 10, 2024

Claims for Patent: 6,632,668

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Summary for Patent: 6,632,668

Title:	Monolayer sheet structure of primary hepatocytes obtained using a protein-phosphorylation inhibitor
Abstract:	A monolayer sheet structure is prepared containing a confluent monolayer of primary hepatocytes cultured on a substrate in sheet form. The monolayer sheet structure is formed by treating the hepatocytes in the monolayer formation stage with a protein-phosphorylation inhibitor having an indolo [2,3-a] carbazole structure, such as staurosporine, or other similar structure.
Inventor(s):	Maeda; Sumio (Aichi, JP), Inagaki; Hidetoshi (Aichi, JP), Saito; Takao (Aichi, JP)
Assignee:	Japan as represented by the Secretary of Agency of Industrial Science and Technology (Tokyo, JP)
Application Number:	09/748,186
Patent Claims:	1. A monolayer sheet structure comprising a confluent monolayer comprising primary hepatocytes cultured on a substrate in sheet form, wherein said monolayer sheet structure of primary hepatocytes is produced by treating the hepatocytes with a protein-phosphorylation inhibitor having an indolo [2,3-a] carbazole skeleton during formation of the monolayer. 2. The monolayer sheet structure according to claim 1, wherein the protein phosphorylation inhibitor is staurosporine. 3. The monolayer sheet structure according to claim 1, wherein the protein-phosphorylation inhibitor is at least one member selected from the group consisting of K252a, K252b, KT5720, and KT5823. 4. The monolayer sheet structure according to claim 1, wherein the protein-phosphorylation inhibitor is at least one member selected from the group consisting of KT5926, UCN-01, UCN-01-Me, RK-286c, and CGP41251. 5. A monolayer sheet structure consisting of a confluent monolayer comprising primary hepatocytes cultured on a substrate in sheet form, wherein said monolayer sheet structure of primary hepatocytes is produced by treating the hepatocytes with a protein-phosphorylation inhibitor having an indolo [2,3-a] carbazole skeleton during formation of the monolayer. 6. A method of manufacturing a monolayer sheet structure comprising culturing primary hepatocytes on a culture substrate in sheet form to produce a confluent monolayer, and treating the hepatocytes with a protein-phosphorylation inhibitor having an indolo [2,3-a] carbazole skeleton during formation of the monolayer to form the monolayer sheet structure of primary hepatocytes. 7. The method according to claim 6, wherein the protein-phosphorylation inhibitor is staurosporine. 8. The method according to claim 6, wherein the protein-phosphorylation inhibitor is at least one member selected from the group consisting of K252a, K252b, KT5720, and KT5823. 9. The method according to claim 6, wherein the protein-phosphorylation inhibitor is at least one member selected from the group consisting of KT5926, UCN-01, UCN-01-Me, RK-286c, and CGP41251. 10. The method according to clam 6, further comprising isolating primary hepatocytes from livers or liver sections by a collagenase perfusion method. 11. The method according to claim 6, further comprising suspending hepatocytes in an animal cell culture medium containing at least one physiologically active hormone. 12. The method according to claim 6, further comprising seeding the hepatocytes on the culture substrate. 13. The method according to claim 6, further comprising subjecting the hepatocytes to a standing culture in the presence of CO.sub.2. 14. The method according to claim 13, wherein the subjecting is performed for a time period up to 24 hours. 15. The method according to claim 13, further comprising exchanging a medium of the standing culture with a second culture medium after 4 hours of subjecting the hepatocytes to the standing culture in the presence of CO.sub.2. 16. The method according to claim 6, further comprising dissolving the protein-phosphorylation inhibitor in an organic solvent. 17. The method according to claim 6, further comprising dissolving the protein-phosphorylation inhibitor in dimethylsulfoxide. 18. The method according to claim 6, wherein the protein-phosphorylation inhibitor is present at a concentration of from 10 to 50 nM.

Details for Patent 6,632,668

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Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965	⤷ Try a Trial	2020-06-07
>Applicant	>Tradename	>Biologic Ingredient	>Dosage Form	>BLA	>Approval Date	>Patent No.	>Expiredate

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