METHODS FOR CULTURING DERMAL CELLS FOR TREATMENT OF SKIN INJURIES SUCH AS BURNS
The present invention relates to novel methods of growing or otherwise producing unpassaged or minimally-passaged dermal cells from a small biopsy specimen for treating skin injuries significantly larger than the biopsy.
1 . A method of generating at least 60 million dermal cells from a small biopsy specimen, comprising the steps of:
a) grinding the small biopsy specimen with a micro tissue grinder to form a ground specimen;
b) digesting the ground specimen with a dissociative or digestive enzyme to form a cell suspension; and
c) harvesting the dermal cells from the cell suspension.
2 . The method of claim 1 , wherein the enzyme is a collagenase.
3 . The method of claim 2 , wherein the collagenase is liberase.
4 . The method of claim 1 , wherein the enzyme is trypsin.
5 . The method of claim 1 , wherein at least 200 million minimally-passaged dermal cells are generated.
6 . A method of generating at least 60 million dermal cells from a small biopsy specimen, comprising the steps of:
a) grinding the small biopsy specimen with a micro tissue grinder to form a ground specimen;
b) digesting the ground specimen with a dissociative or digestive enzyme to form a cell suspension;
c) culturing the cells in the cell suspension for a period from 3 to 10 days; and
d) harvesting the dermal cells.
7 . The method of claim 1 , wherein at least 300 million minimally-passaged dermal cells are generated.
8 . The method of claim 1 , wherein at least 600 million minimally-passaged dermal cells are generated.
9 . The method of claim 1 , wherein the dermal cells have not been passaged.
10 . The method of claim 1 , wherein the dermal cells have been passaged no more than one time.
11 . A method of treating a skin injury in an animal, comprising the step of administering to the animal dermal cells generated by the method of claim 1 .
12 . The method of claim 11 wherein the skin injury is a burn.
13 . The method of claim 11 , wherein one or more administrations each comprises between 10 and 600 million minimally-passaged dermal cells.
14 . The method of claim 12 , wherein the administration is by injection into the burn area or area subadjacent to the burn.
15 . The method of claim 12 , wherein the route of administration is topical application.
16 . A composition comprising minimally passaged dermal cells generated by the method of claim 1 .
17 . A composition comprising conditioned medium produced by the method of claim 6 .