Prevascularized constructs for implantation to provide blood perfusion

Abstract

This application discloses methods and materials for preparing functional microvascular beds in the laboratory. These prevascularized constructs can be used to vascularize engineered tissue constructs or to revascularize damaged or diseased tissues or organs following implantation. The prevascularized constructs may also deliver genetically engineered gene products to the bloodstream.

1 . A method for producing a vascularized tissue, comprising the step of: 
 combining a tissue with a prevascularized construct under conditions and for a time suitable for the vascularization of said tissue.    
   
   
       2 . The method of  claim 1 , wherein said tissue comprises an engineered tissue.  
   
   
       3 . The method of  claim 1 , wherein said tissue is an organ and said vascularization occurs in vivo.  
   
   
       4 . The method of  claim 1 , wherein said construct includes autologous microvessel fragments in a biocompatible three dimensional culture matrix.  
   
   
       5 . The method of  claim 1 , wherein said tissue comprises human cells.  
   
   
       6 . The method of  claim 1 , wherein said prevascularized construct includes cultured microvessel fragments.  
   
   
       7 . The method of  claim 1 , wherein said prevascularized construct comprises human microvessel fragments.  
   
   
       8 . A method for producing a vascularized tissue in a subject, comprising the steps of: 
 (a) combining cells from a selected tissue with a prevascularized construct; and    (b) implanting said combined cells and prevascularized construct into said subject, thereby vascularizing said selected tissue in vivo.    
   
   
       9 . The method of  claim 8 , wherein said selected tissue is chosen from the group consisting of heart tissue, lung tissue, muscle tissue, liver tissue, pancreatic tissue, and lymph tissue.  
   
   
       10 . The method of  claim 8 , wherein said selected tissue is an organ.  
   
   
       11 . The method of  claim 10 , wherein said organ is a liver, heart, lung or other organ suitably transplantable into mammals.  
   
   
       12 . The method of  claim 8 , wherein said construct includes autologous microvessel fragments in a biocompatible three dimensional culture matrix.  
   
   
       13 . The method of  claim 8 , further including the step of genetically modifying said cells with a gene of interest prior to combining said cells with said prevascularized construct.  
   
   
       14 . The method of  claim 8 , wherein said prevascularized construct comprises cultured microvessel fragments.  
   
   
       15 . A method for expanding the vasculature of a diseased tissue in a subject, comprising the step of: 
 combining a prevascularized construct with said diseased tissue in the subject, thereby expanding the vasculature of the diseased tissue in vivo.    
   
   
       16 . The method of  claim 15 , wherein said diseased tissue comprises human tissue.  
   
   
       17 . The method of  claim 16 , wherein said diseased tissue comprises heart tissue.  
   
   
       18 . The method of  claim 15 , wherein said construct includes autologous microvessel fragments in a biocompatible three dimensional culture matrix.  
   
   
       19 . The method of  claim 17 , wherein said construct includes autologous microvessel fragments in a biocompatible three dimensional culture matrix.  
   
   
       20 . The method of  claim 15 , wherein said prevascularized construct further comprises stem cells.  
   
   
       21 . The method of  claim 15 , further including the step of combining genetically modified cells with said prevascularized construct prior to combining said prevascularized construct with said diseased tissue in the subject.  
   
   
       22 . A method for vascularizing a natural tissue in a subject, comprising the steps of: 
 (a) combining a prevascularized construct containing autologous microvessel fragments in a biocompatible three dimensional culture matrix with said natural tissue in vitro; and    (b) implanting said natural tissue into said subject.    
   
   
       23 . The method of  claim 22 , wherein said tissue comprises adipose tissue.  
   
   
       24 . The method of  claim 23 , wherein said adipose tissue is selected from the group consisting of omental fat, properitoneal fat, perirenal fat, pericardial fat, subcutaneous fat, breast fat, or epididymal fat.  
   
   
       25 . The method of  claim 23 , further comprising the step of obtaining said adipose tissue by liposuction, abdominoplasty, or combinations thereof.  
   
   
       26 . The method of  claim 22 , wherein said prevascularized construct further comprises a biocompatible three dimensional culture matrix that includes an autologous fibrin-derived, three-dimensional matrix scaffold.  
   
   
       27 . The method of  claim 22 , wherein at least a portion of said vascularizing said natural tissue occurs in vivo.  
   
   
       28 . The method of  claim 22 , wherein said prevascularized construct further comprises a component selected from the group consisting of collagen, fibrin, gelatin, alginate, hydrogel, dextran, chemically-crosslinkable dextran, or photo-crosslinkable dextran, ePTFE, PTFE, PGA, PLA, PLGA, PET, nylon, vinyl, or combinations thereof.  
   
   
       29 . The method of  claim 22 , wherein said prevascularized construct comprises at least one microvessel fragment obtained from a human and said biocompatible three dimensional culture matrix comprises human collagen, human fibrinogen, human thrombin, or human fibrin.  
   
   
       30 . The method of  claim 22 , wherein said prevascularized construct further comprises a component selected from the group consisting of at least one cytokine, at least one chemokine, at least one antibiotic, at least one drug, at least one analgesic agent, at least one anti-inflammatory agent, at least one immunosuppressive agent, or combinations thereof.  
   
   
       31 . The method of  claim 30 , wherein said at least one cytokine comprises VEGF, PDGF, or both VEGF and PDGF.  
   
   
       32 . A method for expanding microvessel fragments into functional microvessel beds, comprising the steps of: 
 a. isolating said microvessel fragments, and    b. combining the microvessel fragments of step (a) with a biocompatible three dimensional culture matrix under conditions and for a time suitable for the formation said functional microvessel beds.    
   
   
       33 . The method of  claim 32 , wherein said biocompatible three dimensional culture matrix comprises at least one component selected from the group consisting of collagen, fibrin, gelatin, alginate, hydrogel, dextran, chemically-crosslinkable dextran, or photo-crosslinkable dextran, ePTFE, PTFE, PGA, PLA, PLGA, PET, nylon, vinyl, or combinations thereof.