Organ models

Abstract

This document provides materials and methods for making and using functional (e.g., vascularized) organ models (e.g., pancreas models). For example, functional pancreas models including an ECM containing a plurality (e.g., two or more) of pancreatic islets, and a vascular network are provided.

What is claimed is: 
     
         1 . A device capable of being used as a pancreas model, comprising:
 a substrate;   a lower layer of extracellular matrix disposed on top of said substrate;   a strand of sacrificial matrix disposed along an interior region of said lower layer of extracellular matrix;   an array of pancreatic islets disposed on top of said lower layer of extracellular matrix;   an upper layer of extracellular matrix disposed on said array of pancreatic islets; and   a vascular network.   
     
     
         2 . The device of  claim 1 , wherein said lower layer of extracellular matrix comprises fibrin, thrombin, fibrinogen, fibrin hydrogel, collagen, gelatin, a gelatinous protein mixture, and/or laminin. 
     
     
         3 . The device of  claim 1 , wherein said lower layer of extracellular matrix comprises cells selected from the group consisting of endothelial cells, pericytes, fibroblasts, smooth muscle cells, and combinations thereof. 
     
     
         4 . The device of  claim 1 , wherein said lower layer of extracellular matrix comprises calcium chloride. 
     
     
         5 . The device of  claim 1 , wherein said sacrificial matrix is selected from the group consisting of alginate, agarose, gelatin, sugar, and poloxamer. 
     
     
         6 . The device of  claim 1 , wherein said pancreatic islets comprise beta cells and microvessel endothelial cells. 
     
     
         7 . The device of  claim 6 , wherein said beta cells are selected from the group consisting of adipose-derived stem cell derived beta cells, induced pluripotent stem cell (IPS) derived beta cells, fibroblast derived beta cells, and combinations thereof. 
     
     
         8 . The device of  claim 6 , wherein said beta cells and said microvessel endothelial cells are present in a ratio of from about 1:1 to about 10:1. 
     
     
         9 . The device of  claim 1 , wherein said upper layer of extracellular matrix comprises fibrin, thrombin, fibrinogen, fibrin hydrogel, collagen, gelatin, a gelatinous protein mixture, and/or laminin. 
     
     
         10 . The device of  claim 1 , wherein said upper layer of extracellular matrix comprises cells selected from the group consisting of endothelial cells, pericytes, fibroblasts, smooth muscle cells, and combinations thereof. 
     
     
         11 . The device of  claim 1 , wherein said upper layer of extracellular matrix comprises calcium chloride. 
     
     
         12 . A method of making a pancreas model, said method comprising:
 disposing a first layer of extracellular matrix onto a substrate assembled into a model platform;   disposing a strand of sacrificial matrix onto an interior region of said first layer of extracellular matrix;   disposing an array of pancreatic islets onto said first layer of extracellular matrix;   disposing a layer of extracellular matrix onto said array of pancreatic islets;   de-crosslinking said sacrificial matrix with a solution to create a channel;   seeding said channel with microvessel endothelial cells; and   perfusing said channel with perfusate.   
     
     
         13 . The method of  claim 12 , wherein disposing said first layer of extracellular matrix comprises alternately disposing a first layer comprising thrombin, ECs, pericytes, and CaCl 2 , and disposing a second layer comprising fibrinogen on said first layer. 
     
     
         14 . The method of  claim 12 , wherein said sacrificial matrix is selected from the group consisting of alginate, agarose, gelatin, sugar, and poloxamer. 
     
     
         15 . The method of  claim 12 , wherein said pancreatic islets comprise engineered pancreatic spheroids. 
     
     
         16 . The method of  claim 15 , wherein said engineered pancreatic spheroids are engineered by co-culturing β cells and microvessel endothelial cells. 
     
     
         17 . The method of  claim 16 , wherein said beta cells are selected from the group consisting of adipose-derived stem cells beta cells, pluripotent stem cell derived beta cells, fibroblast derived beta cells, and combinations thereof. 
     
     
         18 . The method of  claim 16 , wherein said beta cells and said microvessel endothelial cells are co-cultured in a ratio of from about 1:1 to about 10:1. 
     
     
         19 . The method of  claim 16 , wherein said beta cells and said microvessel endothelial cells are co-cultured in the presence of a growth factor. 
     
     
         20 . The method of  claim 19 , wherein said growth factor is selected from the group consisting of vascular endothelial growth factor, epidermal growth factor, and fibroblast growth factor. 
     
     
         21 . The method of  claim 12 , wherein disposing said second layer of extracellular matrix comprises alternately disposing a first layer comprising thrombin, ECs, pericytes, and CaCl 2 , and disposing a second layer comprising fibrinogen on said first layer until said model platform is full. 
     
     
         22 . The method of  claim 12 , wherein said solution is sodium citrate. 
     
     
         23 . The method of  claim 12 , wherein said channel is perfused with laminar flow. 
     
     
         24 . The method of  claim 12 , wherein said channel is perfused for about 14 days at a rate of about 0.1 dyne/cm 2  for about 6 hours, followed by a rate of about 1 dyne/cm 2  for about 18 hours, followed by a rate of about 10 dyne/cm 2  for about 13 days. 
     
     
         25 . The method of  claim 12 , wherein said perfusate is serum-free media. 
     
     
         26 . The method of  claim 12 , where said pancreas model is a patient-specific pancreas model.