Cultured cartilage/bone cells/tissue, method of generating same and uses thereof

Abstract

A method of generating cultured chondrocytes/endochondral bone cells is provided. The method comprising isolating chondrocytes from mandibular condyle tissue, and culturing the isolated chondrocytes. A method of isolating chondrocytes from mandibular condyle tissue is further provided. The method comprises isolating mandibular condyle tissue from a mammal and treating the mandibular condyle tissue so as to selectively remove fibroblast-like cells and/or myocytes therefrom, the modified mandibular condyle tissue including chondrocytes, and selectively collecting the chondrocytes from the modified mandibular condyle tissue.

1 . A method of generating cultured chondrocytes, the method comprising:
 (a) isolating chondrocytes from mandibular condyle tissue of a neonatal mammal; and   (b) culturing said isolated chondrocytes, wherein said culturing comprises plating said isolated chondrocytes as a monolayer and culturing said isolated chondrocytes for at least 7 days, thereby generating the cultured chondrocytes, wherein the cultured chondrocytes express collagen Type II and not collagen Type I.   
     
     
         2 . The method of  claim 1 , wherein step (a) comprises:
 (c) selectively removing fibroblast-like cells and/or myocytes from said mandibular condyle tissue, thereby generating modified mandibular condyle tissue depleted of said fibroblast-like cells and/or said myocytes, said modified mandibular condyle tissue including chondrocytes; and   (d) selectively isolating said chondrocytes from said modified mandibular condyle tissue.   
     
     
         3 . The method of  claim 2 , wherein step (c) is effected by incubating said mandibular condyle tissue with a protease. 
     
     
         4 . The method of  claim 2 , wherein step (d) is effected by incubating said modified mandibular condyle tissue with a protease so as to selectively release chondrocytes therefrom. 
     
     
         5 . The method of  claim 1 , wherein step (b) is effected in the presence of a culturing medium supplemented with serum, ascorbic acid, β glycerol-phosphate, calcium chloride and pyruvate. 
     
     
         6 . The method of  claim 4 , further comprising isolating said chondrocytes released from said modified mandibular condyle tissue. 
     
     
         7 . The method of  claim 1 , wherein step (b) is effected using culturing conditions including a culture medium devoid of at least one supplement selected from the group consisting of a microfilament-modifying compound, a protein kinase inhibitor, and a polypeptide growth factor, wherein said supplement is not derived from a serum supplement of said culture medium. 
     
     
         8 . The method of  claim 7 , wherein said microfilament-modifying compound is selected from the group consisting of dihydrocytochalasin B, staurosporine, and an actin filament-modifying compound. 
     
     
         9 . The method of  claim 7 , wherein said protein kinase inhibitor is staurosporine and/or a PKC inhibitor. 
     
     
         10 . The method of  claim 7 , wherein said polypeptide growth factor is selected from the group consisting of TGF, FGF, and IGF. 
     
     
         11 . The method of  claim 10 , wherein said TGF is TGF-beta 1. 
     
     
         12 . The method of  claim 10 , wherein said FGF is FGF-2. 
     
     
         13 . The method of  claim 10 , wherein said IGF is IGF-I. 
     
     
         14 . The method of  claim 1 , wherein step (b) is effected using culturing conditions which are normoxic. 
     
     
         15 . The method of  claim 1 , wherein step (b) is effected using culturing conditions which include culturing a subconfluent population of said isolated chondrocytes. 
     
     
         16 . The method of  claim 1 , wherein step (b) is effected for a minimum duration selected from a range of 14-21 days. 
     
     
         17 . A cell culture comprising a monolayer of chondrocytes isolated from a mandibular condyle tissue of a neonatal mammal 
     
     
         18 . The cell culture of  claim 17 , wherein said culture medium includes at least one supplement selected from the group consisting of ascorbic acid, beta-glycerophosphate and pyruvate. 
     
     
         19 . A method of treating a cartilage or bone disease in a subject, the method comprising:
 (a) producing cultured chondrocytes according to the method of  claim 1 ; and   (b) administering a therapeutically effective dose of said cultured chondrocytes to the subject, thereby treating the cartilage or bone disease in the subject.