Claims for Patent: 9,109,204

Summary for Patent: 9,109,204

Title:	Methods for compact aggregation of dermal cells
Abstract:	The invention provides for a method for aggregating dermal papilla cells or dermal sheath cells or a combination thereof, the method comprising: growing dermal papilla cells or dermal sheath cells or a combination thereof in suspension culture; and contacting the culture with an effective amount of an enzyme, wherein a substrate of the enzyme is an extracellular matrix molecule in the suspension culture, so as to aggregate dermal papilla cells or dermal sheath cells. The culture may be a hanging drop culture and the enzyme may be a hyaluronidase.
Inventor(s):	Christiano; Angela M. (Upper Saddle River, NJ), Jahoda; Colin A. B. (Durham, GB)
Assignee:	THE TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK (New York, NY) DURHAM UNIVERSITY (Durham, GB)
Application Number:	12/280,661
Patent Claims:	1. A method for maintaining hair inductive activity of dermal papilla cells or dermal sheath cells or a combination thereof, the method comprising: growing dermal papilla cells or dermal sheath cells or a combination thereof in suspension culture so as to obtain a compact aggregate of cells, wherein the suspension culture is a hanging drop culture. 2. The method of claim 1, further comprising: admixing with the suspension culture an effective amount of a substance capable of reducing the amount of extracellular matrix in the suspension culture. 3. The method of claim 1 or 2, wherein the suspension culture comprises a soluble factor. 4. The method of claim 3, wherein the soluble factor is added exogenously. 5. The method of claim 3, wherein the soluble factor is administered in an amount of from about 5 ng/ml of culture media to about 300 ng/ml of culture media. 6. The method of claim 3, wherein the soluble factor comprises periostin, follistatin, Wise, Wnt10b, any one or more soluble factors in Table 1 and Table 2, or any combination thereof. 7. The method of claim 2, wherein the substance is a protein. 8. The method of claim 7, wherein the protein is an enzyme. 9. The method of claim 8, wherein a substrate of the enzyme is an extracellular matrix molecule in the suspension culture. 10. The method of claim 8, wherein the enzyme is a hyaluronidase, a collagenase, a chondroitinase, or a combination thereof. 11. The method of claim 10, wherein the hyaluronidase is admixed with the suspension culture in an amount of from about 20 U/ml of culture media to about 50 U/ml of culture media for up to about 15 days. 12. The method of claim 10, wherein the hyaluronidase is Hyal-1, Hyal-2, Hyal-3, or a combination thereof. 13. The method of claim 7, wherein the protein degrades one or more molecules in the extracellular matrix. 14. The method of claim 1, wherein the hanging drop culture contains less than about 9,000 cells. 15. The method of claim 1, wherein the hanging drop culture contains less than about 7,000 cells. 16. The method of claim 1, wherein the hanging drop culture contains less than about 5,000 cells. 17. The method of claim 1, wherein the hanging drop culture contains less than about 3,000 cells. 18. The method of claim 1, wherein the suspension culture further comprises epithelial cells. 19. The method of claim 18, wherein the epithelial cells are derived from hair follicle or skin. 20. The method of claim 18, wherein the epithelial cell is a keratinocyte. 21. The method of claim 1, wherein the hanging drop is cultured for at least 24 hours. 22. The method of claim 1, wherein the hanging drop is cultured for at least 48 hours. 23. The method of claim 1, wherein the hanging drop is cultured up to about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days. 24. The method of claim 1, wherein the hanging drop is cultured until expression of an inductivity marker gene is reduced. 25. The method of claim 24, wherein the inductivity marker gene comprises Wnt10b, WISE, versican, or a combination thereof. 26. The method of claim 1 or 2, further comprising grafting the compact aggregate of cells into the skin of a subject. 27. The method of claim 26, wherein the subject is a mammal. 28. The method of claim 27, wherein the mammal is a human, a mouse, a dog, a cat, a horse, a cow, or a bird. 29. The method of claim 26, wherein the cells are autologous to the subject. 30. The method of claim 1 or 2, wherein the cells comprise primary cells, secondary cells, passaged secondary cells, or a cell line. 31. The method of claim 30, wherein the cells are obtained or derived from a mammal. 32. The method of claim 31, wherein the mammal is a human, a mouse, a dog, a cat, a horse, or a cow. 33. The method of claim 1, further comprising promoting formation of hair follicles from the aggregated dermal papilla cells or dermal sheath cells or a combination thereof, the method comprising: admixing hyaluronidase with the culture for a time of from about 1 day to about 15 days; and growing the culture for a sufficient time so as form hair follicles. 34. The method of claim 33, wherein the suspension culture comprises a soluble factor. 35. The method of claim 34, wherein the soluble factor is added to the growing suspension culture. 36. The method of claim 34, wherein the soluble factor is admixed to the suspension culture in an amount of from about 5 ng/ml of culture media to about 300 ng/ml of culture media. 37. The method of claim 34, wherein the soluble factor comprises periostin, follistatin, Wise, Wnt10b, any one or more soluble factors in Table 1 and Table 2, or any combination thereof. 38. The method of claim 33, wherein the hanging drop culture contains less than about 9,000 cells. 39. The method of claim 33, wherein the hanging drop culture contains less than about 7,000 cells. 40. The method of claim 33, wherein the hanging drop culture contains less than about 5,000 cells. 41. The method of claim 33, wherein the hanging drop culture contains less than about 3,000 cells. 42. The method of claim 33, wherein the suspension culture further comprises epithelial cells. 43. The method of claim 42, wherein the epithelial cells are derived from a hair follicle or skin. 44. The method of claim 42, wherein the epithelial cell is a keratinocyte. 45. The method of claim 33, wherein the hyaluronidase is admixed in an amount of from about 20 U/ml of culture media to about 50 U/ml of culture media. 46. The method of claim 33, wherein the hyaluronidase is Hyal-1, Hyal-2, Hyal-3, or a combination thereof. 47. The method of claim 33, wherein the suspension culture is grown for at least 24 hours. 48. The method of claim 33, wherein the suspension culture is cultured until expression of an inductivity marker gene is reduced. 49. The method of claim 48, wherein the inductivity marker gene comprises Wnt10b, WISE, versican, or a combination thereof. 50. The method of claim 33, further comprising grafting the hair follicles into the skin of a subject. 51. The method of claim 50, wherein the subject is a mammal. 52. The method of claim 50, wherein the cells are autologous to the subject. 53. The method of claim 51, wherein the mammal is a human, mouse, dog, or cat. 54. The method of claim 33, wherein the cells comprise primary cells, secondary cells, passaged secondary cells, or a cell line. 55. The method of claim 54, wherein the cells are from a mammal. 56. The method of claim 55, wherein the mammal is a human, a mouse, a dog, a cat, a horse, or a cow.

Details for Patent 9,109,204

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Bausch & Lomb Incorporated	VITRASE	hyaluronidase	Injection	021640	05/05/2004		2026-02-28
Bausch & Lomb Incorporated	VITRASE	hyaluronidase	Injection	021640	12/02/2004		2026-02-28
Amphastar Pharmaceuticals, Inc.	AMPHADASE	hyaluronidase	Injection	021665	10/26/2004		2026-02-28
Akorn, Inc.	HYDASE	hyaluronidase	Injection	021716	10/25/2005		2026-02-28
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2026-02-28