Decellularized tissue engineered constructs and tissues

Abstract

Methods for producing tissue engineered constructs and engineered native tissues include producing a tissue engineered construct by growing cells in vitro on a substrate and then decellularizing the construct to produce a decellularized construct consisting largely of extracellular matrix components. The construct can be used immediately or stored until needed. The decellularized construct can be used for further tissue engineering, which may include seeding the construct with cells obtained from the intended recipient of the construct. During any of the growth phases required for production of the construct, the developing construct may be subjected to various tissue engineering steps such as application of mechanical stimuli including pulsatile forces.

1 . A decellularized construct for use as a medicament for implanting into a subject comprising:
 a decellularized proteinaceous extracellular matrix having a thickness of greater than 50 μm, wherein the matrix was formed by culturing a plurality of individual cells of a first type in a three dimensional mass of living tissue on an artificial substrate comprising a synthetic polymeric material.   
     
     
         2 . The construct of  claim 1 , wherein at least 80% of the cells of the first type are absent from the construct. 
     
     
         3 . The construct of  claim 1 , wherein at least 90% of the cells of the first type are absent from the construct. 
     
     
         4 . The construct of  claim 1 , wherein at least 95% of the cells of the first type are absent from the construct. 
     
     
         5 . The construct of  claim 1 , wherein at least 99% of the cells of the first type are absent from the construct. 
     
     
         6 . The construct of  claim 1 , wherein at most 10% of the cells of the first type are present in the construct. 
     
     
         7 . The construct of  claim 1 , wherein at most 5% of the cells of the first type are present in the construct. 
     
     
         8 . The construct of  claim 1 , wherein at most 1% of the cells of the first type are present in the construct. 
     
     
         9 . The construct of  claim 1 , further comprising living cells of a second type seeded on the proteinaceous extracellular matrix. 
     
     
         10 . The construct of  claim 9 , wherein said second type of cells are derived from the subject. 
     
     
         11 . The construct of  claim 1  wherein the individual cells are smooth muscle cells. 
     
     
         12 . The construct of  claim 1  wherein the living cells are allogeneic to the subject. 
     
     
         13 . A construct for use in the manufacture of a medicament for implanting into a subject comprising: a freeze-thawed, proteinaceous extracellular matrix, wherein the proteinaceous extracellular matrix has a thickness of greater than 50 μm, and wherein the proteinaceous extracellular matrix was formed around a plurality of individual cells which were cultured in a three dimensional mass of living tissue on a flat or tubular artificial substrate. 
     
     
         14 . The construct of  claim 13  wherein the individual cells are smooth muscle cells. 
     
     
         15 . The construct of  claim 13  wherein the living cells are allogeneic to the subject.