Claims for Patent: 8,728,819

Summary for Patent: 8,728,819

Title:	Methods for culturing minimally-passaged fibroblasts and uses thereof
Abstract:	The present invention relates to novel methods of growing at least 100 million minimally-passaged fibroblasts from a small biopsy specimen. The invention includes methods wherein a small biopsy specimen is seeded directly into a large tissue culture flask, and passaged no more than three times.
Inventor(s):	Maslowski; John (Pottstown, PA), Thomas; Myrna F. (Hainesport, NJ), Lindner; Marie A. (Radnor, PA)
Assignee:	Fibrocell Technologies, Inc. (Exton, PA)
Application Number:	12/438,932
Patent Claims:	1. A cell-free topical composition comprising derived from at most passage three fibroblasts conditioned medium produced by a method comprising the steps of: a) digesting a tissue biopsy specimen with a dissociative or digestive enzyme to form a fibroblast cell suspension, wherein the biopsy specimen is a single full-thickness biopsy not more than 5 mm thick and not more than 12 cm.sup.2; b) seeding the fibroblast cell suspension into a first tissue culture flask containing a medium; c) culturing a fibroblast cell population in the first tissue culture flask through at most three passages into sequentially larger tissue culture flasks in the medium, thereby generating at least 100 million minimally-passaged fibroblasts and a conditioned medium; d) harvesting said at least 100 million minimally-passaged fibroblasts to obtain a cell-free conditioned medium; and e) formulating the conditioned medium for topical administration to obtain the cell-free topical composition. 2. The composition of claim 1, wherein the enzyme is a collagenase. 3. The composition of claim 2, wherein the collagenase is liberase. 4. The composition of claim 1, wherein the enzyme is trypsin. 5. The composition of claim 1, wherein at least 200 million minimally-passaged fibroblasts are generated. 6. The composition of claim 1, wherein at least one billion minimally-passaged fibroblasts are generated. 7. The composition of claim 1, wherein the first tissue culture flask is a T-175 or a T-225 flask. 8. The composition of claim 7, wherein the sequentially larger tissue culture flasks are a T-500 flask or a 10-layer cell culture stack. 9. The composition of claim 1, wherein the first tissue culture flask is a T-175 or a T-225 flask, the sequentially larger tissue culture flasks are a T-500 flask or multilayer cell culture stacks cell culture stack, and wherein the multilayer stacks are 10-layer stacks. 10. The composition of claim 1, wherein the at least 100 million minimally-passaged fibroblasts are generated by culturing the fibroblast cell population in the first tissue culture flask through no more than three passages into sequentially larger tissue culture flasks.

Details for Patent 8,728,819

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2039-03-29