Autogenic Living Scaffolds and Living Tissue Matrices: Methods and Uses Thereof

Abstract

The present invention is drawn to a 3-dimensional cell-produced scaffold construct comprising cells and the extracellular matrix that has been produced and arranged by these cells.

1 - 91 . (canceled) 
     
     
         92 . A three-dimensional cell-produced scaffold construct comprising cells and extracellular matrix synthesized by said cells, wherein the scaffold construct has:
 a thickness of greater than 100 μm;   a porosity that enables cellular migration of additional cells throughout the construct; and   
       wherein the extracellular matrix is synthesized in vitro by said cells. 
     
     
         93 . A three-dimensional cell-produced scaffold construct comprising extracellular matrix synthesized in vitro by cells, wherein said construct is produced by a process comprising:
 i) incubating the cells at hyperconfluent density in defined chemical media which causes the cells to enter a synthetic phase that promotes extracellular matrix synthesis of said cells without significant proliferation; and   ii) isolating the construct from the media,   
       wherein the cells are incubated for a period of time in the media that is sufficient to produce a construct having a thickness and porosity that enables cellular migration of additional cells throughout the construct. 
     
     
         94 . A three-dimensional cell-produced scaffold construct comprising cells and extracellular matrix synthesized in vitro by said cells, wherein said construct has a thickness and porosity that enables cellular migration throughout the construct, wherein the construct has a growth rate in the media such that it is capable of achieving a thickness of greater than 100 μm, wherein the construct is made by a process comprising:
 i) incubating the cells at hyperconfluent density in defined chemical media which causes the cells to enter a synthetic phase that promotes extracellular matrix synthesis of said cells without significant proliferation; and 
 ii) isolating the construct from the media. 
 
     
     
         95 . A three-dimensional cell-produced scaffold construct comprising extracellular matrix synthesized in vitro by cells, wherein said construct is produced by a process comprising:
 i) incubating the cells at hyperconfluent density in defined chemical media which causes the cells to enter a synthetic phase that promotes extracellular matrix synthesis of said cells without significant proliferation; and   ii) isolating the construct from the media,   
       wherein the construct has a porosity such that it is capable of achieving a thickness of greater than 100 μm. 
     
     
         96 . The construct of  claim 93 , wherein the construct has a growth rate in the media such that it is capable of achieving a thickness of greater than 100 μm within three weeks of growth. 
     
     
         97 . The construct of  claim 92 , wherein the cells are seeded at a density of between 200,000 to 1,000,000 cells/cm 2 . 
     
     
         98 . The construct of  claim 92 , wherein said cells are maintained at a density of about 1,000 cells/mm 3  to greater than about 200,000 cells/mm 3 . 
     
     
         99 . The construct of  claim 92 , wherein some of said cells are genetically engineered. 
     
     
         100 . The construct of  claim 92 , wherein said construct promotes differentiation, dedifferentiation, transdifferentiation or any combination thereof, of cells or tissue either within or in contact with the construct in vitro or in vivo. 
     
     
         101 . The construct of  claim 92 , wherein said construct promotes cell growth, proliferation, migration and/or acquisition of in vivo-like morphology or any combination thereof, either within or in contact with the construct in vitro or in vivo. 
     
     
         102 . The construct of  claim 92 , wherein said construct is an ex vivo cell-produced tissue. 
     
     
         103 . The construct of  claim 92 , wherein said construct provides mechanical support, structural support, nutritional support, developmental support, support for wound repair, or supports the growth or regeneration of cells, tissue, organs, or any combination thereof. 
     
     
         104 . The construct of  claim 92 , wherein said construct begins attaching to a site of implantation within about 36 hours, and continues attaching for several days, weeks, or months depending on the site of implantation and degree of immune system activity at the site of implantation. 
     
     
         105 . The construct of  claim 92 , wherein said construct further comprises additional cells, tissue or any combination thereof.