Perfusion bioreactor

Abstract

In some embodiments the present invention provides perfusion bioreactors and cell culture scaffolds suitable for the preparation of tissue grafts, such as bone tissue grafts. In some embodiments, the perfusion bioreactors comprise a graft chamber and/or a graft chamber insert configured to hold a tissue graft having a certain shape and/or certain dimensions, and/or to allow culture of such tissue grafts under press-fit direct perfusion conditions. In some embodiments, the perfusion bioreactors comprise an equilibration chamber.

What is claimed is: 
     
         1 . A perfusion bioreactor suitable for use in the preparation of a tissue graft segment comprising:
 a) a graft chamber; and   b) an equilibration chamber in fluid communication with the graft chamber.   
     
     
         2 . The perfusion bioreactor of  claim 1 , wherein the graft chamber is configured to accommodate a tissue graft segment. 
     
     
         3 . The perfusion bioreactor of  claim 1 , wherein the graft chamber further comprises a graft chamber insert configured to accommodate a tissue graft segment. 
     
     
         4 . The perfusion bioreactor of  claim 1 , comprising a bottom portion and a top portion, the bottom portion comprising: i) the graft chamber, ii) the equilibration chamber, iii) an inlet, and iv) a fluid channel defining a fluid path between the inlet and the equilibration chamber. 
     
     
         5 . The perfusion bioreactor of  claim 4 , wherein the top portion comprises: i) a fluid reservoir, ii) an aperture fluidly connecting the fluid reservoir and the graft chamber, and iii) an outlet port. 
     
     
         6 . The perfusion bioreactor of  claim 5 , wherein the bottom and top portions are attached via a fastening mechanism. 
     
     
         7 . The perfusion bioreactor of  claim 1 , further comprising an inlet, a fluid channel defining a fluid path between the inlet and the equilibration chamber, a fluid reservoir, an aperture fluidly connecting the fluid reservoir and the graft chamber, the fluid reservoir further comprising an outlet port. 
     
     
         8 . The perfusion bioreactor of  claim 7 , wherein the graft chamber is disposed adjacent the equilibration chamber. 
     
     
         9 . The perfusion bioreactor of  claim 8 , further comprising a fluid pump and one or more tubes fluidly connecting the inlet port and the outlet port to the pump thereby providing a fluid circuit. 
     
     
         10 . The perfusion bioreactor of  claim 1 , wherein the graft chamber is custom-designed to accommodate the tissue graft segment. 
     
     
         11 . The perfusion bioreactor of  claim 3 , wherein the graft chamber insert is custom-designed to accommodate the tissue graft segment. 
     
     
         12 . The perfusion bioreactor of  claim 1 , wherein the graft chamber is custom-designed to accommodate the tissue graft segment using a digital three-dimensional model of the tissue graft segment. 
     
     
         13 . The perfusion bioreactor of  claim 3 , wherein the graft chamber insert is custom-designed to accommodate the tissue graft segment using a digital three-dimensional model of the tissue graft segment. 
     
     
         14 . The perfusion bioreactor of  claim 1 , wherein the tissue graft segment has a maximum thickness of about one centimeter or less. 
     
     
         15 . The perfusion bioreactor of  claim 1 , wherein the tissue graft segment has a maximum thickness of about 0.3 millimeters to 10 millimeters. 
     
     
         16 . The perfusion bioreactor of  claim 12 , wherein the digital three-dimensional model of the tissue graft segment is generated by medical imaging, computed tomography, computer-assisted design, or any combination thereof. 
     
     
         17 . The perfusion bioreactor of  claim 13 , wherein the digital three-dimensional model of the tissue graft segment is generated by medical imaging, computed tomography, computer-assisted design, or any combination thereof. 
     
     
         18 . The perfusion bioreactor of  claim 1 , wherein the equilibration chamber comprises a flat floor. 
     
     
         19 . The perfusion bioreactor of  claim 1 , wherein the equilibration chamber comprises a tapered floor. 
     
     
         20 . The perfusion bioreactor of  claim 19 , wherein the taper forms an angle of about 10, 20, 30, 40, 50, 60 or 70 degrees between the floor and the equilibration chamber wall. 
     
     
         21 . The perfusion bioreactor of  claim 7 , wherein the equilibration chamber further comprises one or more diffusion enhancing elements. 
     
     
         22 . The perfusion bioreactor of  claim 7 , wherein the equilibration chamber further comprises an insert to maintain the dimensions of the equilibration chamber and/or maintain fluid flow through the perfusion bioreactor. 
     
     
         23 . The perfusion bioreactor of  claim 7 , wherein the graft chamber further comprises an insert to (i) maintain the size and dimensions of the graft chamber and equilibrium chamber, and/or (ii) maintain fluid flow through the perfusion bioreactor. 
     
     
         24 . The perfusion bioreactor of  claim 6 , wherein the fastening mechanism comprises screws, rods, pins, clips, latches or any combination thereof. 
     
     
         25 . The perfusion bioreactor of  claim 6 , further comprising an o-ring or other sealing device capable of preventing fluid leakage from between the bottom and top portions. 
     
     
         26 . The perfusion bioreactor of  claim 9 , wherein the pump is a peristaltic pump. 
     
     
         27 . The perfusion bioreactor of  claim 6 , further comprising a sealing mechanism situated between the top and the bottom portions. 
     
     
         28 . The perfusion bioreactor of  claim 4 , wherein the bottom portion and top portion are formed as a single unitary piece. 
     
     
         29 . The perfusion bioreactor of  claim 6 , wherein the top and bottom portions further comprise one or more holes to facilitate the fastening mechanism.