Tissue Repair Using Allogenic Dermal Fibroblasts

Abstract

The present invention provides a method for the repair of subcutaneous or dermal tissue in a subject, comprising in one aspect the injection of a suspension of allogeneic dermal fibroblasts into the subject. By injecting a suspension of allogeneic cells, the invention provides, for example, long-term augmentation of the subadjacent tissue without the disadvantages that accompany the preparation and/or use of presently available materials such as autologous fibroblasts cells or collagen. According to a first aspect of the invention there is provided a method for the augmentation of subcutaneous or dermal tissue in a subject, which method comprises the steps of: (i) providing a suspension of allogeneic dermal fibroblasts; and (ii) injecting an effective volume of the suspension into tissue subadjacent to the subcutaneous or dermal tissue so that the tissue is augmented. The method is preferably cosmetic.

1 - 20 . (canceled) 
     
     
         21 . A method for the augmentation of subcutaneous or dermal tissue in a subject, which method comprises the steps of:
 (i) providing a suspension of allogeneic dermal fibroblasts; and   (ii) injecting an effective volume of the suspension into tissue subadjacent to the subcutaneous or dermal tissue so that the tissue is augmented.   
     
     
         22 . The method of  claim 21 , wherein the method is cosmetic. 
     
     
         23 . The method of  claim 21 , further comprising a step of identifying a defect that is susceptible to amelioration by augmentation of the subadjacent subcutaneous or dermal tissue. 
     
     
         24 . The method of  claim 21 , in which the fibroblasts are passaged. 
     
     
         25 . The method of  claim 23 , wherein the defect is a rhytid, stretch mark, a depressed scar, a cutaneous depression of non-traumatic origin or an underdevelopment of the lip. 
     
     
         26 . The method of  claim 21 , wherein the suspension further comprises fibrin. 
     
     
         27 . The method of  claim 26 , wherein the fibrin is at a final concentration of 0.1 to 20 mg/ml. 
     
     
         28 . The method of  claim 26 , wherein the fibrin is human fibrin. 
     
     
         29 . The method of  claim 21 , wherein the suspension further comprises collagen or hyaluronic acid. 
     
     
         30 . The method of  claim 21 , which further comprises the steps of:
 a) obtaining a dermal biopsy from a source allogeneic to the subject;   b) passaging the dermal fibroblasts from the dermal biopsy in a culture medium comprising between 0.5% and 20% non-human serum, so as to provide dermal fibroblasts substantially free of adipocytes, keratinocytes and extracellular matrix; and   c) forming a suspension of the incubated fibroblasts.   
     
     
         31 . The method of  claim 30 , in which the dermal biopsy is from human neonatal foreskin. 
     
     
         32 . The method of  claim 30 , in which the suspension is formed by scraping the fibroblasts and/or exposing the fibroblasts to a proteolytic enzyme and/or an animal product free nonenzymatic alternative. 
     
     
         33 . The method of  claim 21 , in which the subject is human. 
     
     
         34 . The method of  claim 21 , wherein between 10 4  and 10 8  fibroblasts are injected into said subject. 
     
     
         35 . The method of  claim 34 , wherein between 10 6  and 10 8  fibroblasts are injected into said subject. 
     
     
         36 . The method of  claim 34 , wherein 1 to 5×10 7  fibroblasts are injected into said subject. 
     
     
         37 . The method of  claim 34 , wherein 1×10 7  fibroblasts are injected into said subject. 
     
     
         38 . The method of  claim 21 , wherein a volume of about 1 ml of suspension is injected. 
     
     
         39 . The method of  claim 21 , wherein the cells are suspended in a preservation medium. 
     
     
         40 . The method of  claim 39 , wherein the fibroblasts are suspended in the preservation medium at a temperature of about 2° C. to about 8° C. for at least one hour. 
     
     
         41 . A method of treating subcutaneous or dermal tissue defects by administering a composition comprising allogeneic dermal fibroblasts, wherein said composition optionally includes a preservation medium. 
     
     
         42 . A method of cosmetically treating subcutaneous or dermal tissue defects by administering a composition comprising allogeneic dermal fibroblasts, wherein said composition optionally includes a preservation medium. 
     
     
         43 . A composition for the augmentation of subcutaneous or dermal tissue in a subject, which comprises a suspension of allogeneic dermal fibroblasts in a preservation medium.