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.

1 - 20 . (canceled) 
     
     
         21 . A method of treatment of a defect in skin, tissue, or bone in a human, the method comprising:
 administering to the defect a dose of a pharmaceutical composition comprising at least 10-20 million viable minimally passaged fibroblasts; and   assessing if the defect is repaired to a desired result.   
     
     
         22 . The method of  claim 21 , wherein the administering is in an area subadjacent to the defect. 
     
     
         23 . The method of  claim 21 , wherein the defect is in skin. 
     
     
         24 . The method of  claim 21 , wherein the defect is in tissue. 
     
     
         25 . The method of  claim 21 , wherein the defect is in bone. 
     
     
         26 . The method of  claim 21 , wherein the defect is selected from the group consisting of rhytids, stretch marks, depressed scars, cutaneous depressions of non-traumatic origin, scarring from acne vulgaris, hypoplasia of the lip, vocal cord defects, defects of the oral mucosa, the gingival mucosa, or the palatal mucosa or skin, burns, and burn scars. 
     
     
         27 . The method of  claim 21 , wherein the composition is administered topically. 
     
     
         28 . The method of  claim 21 , wherein the composition is administered via injection. 
     
     
         29 . The method of  claim 24 , wherein the desired result is the regeneration of the tissue. 
     
     
         30 . The method of  claim 24 , wherein the desired result is repair of the tissue defect. 
     
     
         31 . The method of  claim 23 , wherein the desired result is augmentation of the skin. 
     
     
         32 . The method of  claim 21 , wherein the desired result is without surgery. 
     
     
         33 . The method of  claim 21 , wherein the fibroblasts are autologous fibroblasts. 
     
     
         34 . The method of  claim 21 , wherein the composition further comprises acellular matrices. 
     
     
         35 . The method of  claim 21 , wherein the composition further comprises basic fibroblast growth factor. 
     
     
         36 . The method of  claim 21 , the method further comprising repeating administering the dose of the pharmaceutical composition comprising at least 10-20 million viable minimally passaged fibroblasts to the human. 
     
     
         37 . A method of treatment of a tissue defect in a human, the method comprising:
 injecting into the tissue defect a dose of a pharmaceutical composition comprising at least 10-20 million viable minimally passaged fibroblasts; and   assessing if the defect is repaired to a desired result.   
     
     
         38 . The method of  claim 37 , wherein the composition comprises acellular matrices. 
     
     
         39 . The method of  claim 37 , wherein the desired result is augmentation of the tissue defect. 
     
     
         40 . A method of treatment of a tissue defect in a human, the method comprising:
 topically administering to the tissue defect a dose of a pharmaceutical composition comprising at least 10-20 million viable minimally passaged fibroblasts; and   assessing if the defect is repaired to a desired result, without surgery.