Three-dimensional, prevascularized, engineered tissue constructs, methods of making and methods of using the tissue constructs

Abstract

Three-dimensional (3D), prevascularized, engineered tissue constructs, 3D prevascularized engineered tissue models of cancer, and bioreactors and bioreactor arrays including the tissue constructs are disclosed. Methods of making the tissue constructs, methods of using the tissue constructs, methods of drug discovery using the tissue constructs and/or cancer models, and the like are also disclosed.

1 . A three-dimensional (3D), engineered, vascularized tissue construct comprising:
 a 3D, biocompatible scaffold material comprising a solid, porous material and a gel matrix material, wherein the solid, porous material is chosen from the group consisting of: aerogels, reticulate vitreous carbon (RVC), particle stabilized foam, electrospun polymers, synthetic electrospun polymers, and combinations thereof, and wherein the gel matrix material is chosen from the group consisting of: synthetic hydrogels, naturally-derived hydrogels, and a combination thereof;   a 3D network of living cells within the scaffold material;   a plurality of microchannels extending through the construct, such that a substantial portion of the channels have an inlet at one surface of the construct and an outlet at an opposing surface of the construct, wherein the channels form a lumen for allowing passage of fluid through the construct; and   a plurality of endothelial cells at least partially lining the lumen,   
       wherein the endothelial cells result from cells seeded within the lumen of the microchannel and the 3D network of living cells in the scaffold material results from cells seeded within the scaffold material. 
     
     
         2 . The 3D vascularized tissue construct of  claim 1 , wherein the gel matrix material further comprises a crosslinking agent, a gelling agent, or a combination thereof that increases the structural stability of the gel matrix material. 
     
     
         3 . The 3D vascularized tissue construct of  claim 1 , wherein the synthetic hydrogel is selected from the group consisting of: alginate, biocompatible polymer hydrogels, biocompatible copolymer hydrogels, polyethylene glycol (PEG) based hydrogels, and combinations thereof, and wherein the naturally-derived hydrogel is selected from the group consisting of: collagen, fibrin, elastin, keratin, bacterial cellulose, animal-derived basement membrane extract, and combinations thereof. 
     
     
         4 . The 3D vascularized tissue construct of  claim 1 , wherein the biocompatible scaffold material comprises a solid porous material and a combination of at least two gel matrix materials. 
     
     
         5 . The 3D vascularized tissue construct of  claim 4 , wherein the gel matrix material comprises a combination of collagen and an animal-derived basement membrane extract. 
     
     
         6 . The 3D vascularized tissue construct of  claim 1 , wherein the tissue construct is configured to be placed in a well-plate or a flattened parallel plate flow chamber. 
     
     
         7 . The 3D vascularized tissue construct of  claim 1 , wherein the tissue construct is configured for placement on a confocal imaging device, a fluorescence imaging device, a phase contrast imaging device, or interchangeably on any of these devices. 
     
     
         8 . The 3D vascularized tissue construct of  claim 1 , wherein the microchannels have a diameter of about 450 microns or less. 
     
     
         9 . The 3D vascularized tissue construct of  claim 1 , wherein the 3D network of cells comprise mammalian cells, wherein the cells are pathologic or healthy mammalian cells or a combination thereof. 
     
     
         10 . The 3D vascularized tissue construct of  claim 1 , wherein the 3D network of cells comprise cancer cells or a combination of cancer cells and non-cancerous cells. 
     
     
         11 . The 3D vascularized tissue construct of  claim 10 , wherein the non-cancerous comprise fibroblasts, epithelial cells, human tissue cells, or a combination thereof. 
     
     
         12 . A three-dimensional (3D) vascularized biocompatible scaffold for supporting in vitro, 3D tissue culture comprising:
 a 3D, biocompatible scaffold material comprising a solid, porous material and a gel matrix material, wherein the solid, porous material is chosen from the group consisting of: aerogels, reticulate vitreous carbon, particle stabilized foam, and combinations thereof, and wherein the gel matrix material is chosen from the group of gel matrix materials consisting of: synthetic hydrogels, naturally-derived hydrogels, and a combination thereof; and   a plurality of channels extending through the scaffold, such that a substantial portion of the channels have an inlet at one surface of the scaffold and an outlet at an opposing surface of the scaffold, wherein the channels form lumen for allowing passage of liquid through the scaffold.   
     
     
         13 . The 3D vascularized biocompatible scaffold of  claim 12 , further comprising a plurality of endothelial cells at least partially lining the lumen. 
     
     
         14 . A method of making a three-dimensional (3D), vascularized, tissue construct in vitro comprising:
 providing a 3D vascularized biocompatible scaffold, wherein the scaffold comprises:
 a 3D, biocompatible scaffold material comprising a solid, porous material, a gel matrix material, or a combination thereof, wherein the solid, porous material is chosen from the group consisting of: aerogels, reticulate vitreous carbon, and particle stabilized foam, and wherein the gel matrix material is chosen from the group consisting of: synthetic hydrogels, naturally-derived hydrogels, and a combination thereof; and 
 a plurality of channels extending through the scaffold, such that a substantial portion of the channels have an inlet at one surface of the scaffold and an outlet at an opposing surface of the scaffold, wherein the channels form lumen for allowing passage of liquid through the scaffold; 
   seeding the scaffold with cells of at least one tissue type;   perfusing the scaffold with cell culture media; and   incubating the seeded scaffold,   
       wherein a 3D network of cells of the at least one tissue type grows within the scaffold. 
     
     
         15 . The method of  claim 14 , further comprising seeding the lumen with endothelial cells, such that endothelial cells line at least a portion of the lumen. 
     
     
         16 . The method of  claim 14 , wherein the plurality of microchannels are formed in the scaffold material with a microchannel construct comprising a plurality of rods or wires for forming the channels. 
     
     
         17 . The method of  claim 16 , wherein the microchannel construct is inserted into the scaffold material before the gel matrix material is cured and is removed from the scaffold material after the gel matrix material is cured. 
     
     
         18 . A perfusion bioreactor comprising:
 a three-dimensional (3D), engineered tissue construct comprising:
 a 3D, biocompatible scaffold material comprising a solid, porous material and a gel matrix material, wherein the solid, porous material is chosen from the group consisting of: aerogels, reticulate vitreous carbon, and particle stabilized foam, and wherein the gel matrix material is chosen from the group consisting of: synthetic hydrogels, naturally-derived hydrogels, and a combination thereof; 
 a 3D network of living cells within the scaffold material; 
 a plurality of channels extending through the construct, such that a substantial portion of the channels have an inlet at one surface of the construct and an outlet at an opposing surface of the construct, wherein the channels form lumen for allowing passage of fluid media through the construct; and 
 a plurality of endothelial cells at least partially lining the lumen; 
   a tissue chamber configured to house the tissue construct such that the tissue construct forms a barrier between upstream and downstream flow of media through the chamber directing flow of media through the channels of the tissue construct, wherein the chamber has at least one input portal upstream of the location of the tissue construct and at least one output portal downstream of the location of the tissue construct; and   at least one pump to control flow of media through the tissue chamber.   
     
     
         19 . The perfusion bioreactor of  claim 18 , wherein the at least one pump is selected from the group consisting of: micropumps, syringe pumps, peristaltic pumps, and combinations thereof. 
     
     
         20 . The perfusion bioreactor of  claim 18 , further comprising:
 one or more additional perfusion bioreactors, wherein the perfusion bioreactors are interconnected to form a bioreactor network comprising an array of interconnected bioreactors; and   at least one pump to control flow of media through the bioreactors, wherein the bioreactors in the array are in fluid communication with each other, and   
       wherein at least one bioreactor in the array contains cells from a different tissue type than the cells in at least one other bioreactor in the array.