Patent ID: 8728819

Claim:
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 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.