Device and Method of 3-Dimensionally Generating IN VITRO Blood Vessels

Abstract

Provided herein are apparatuses, systems and methods for generating concentration gradients of soluble molecules. Also, provided herein devices and methods for generating in vitro blood vessels.

1 . A blood vessel-generating device comprising:
 a) a sink channel in fluid communication with a sink inlet;   b) a source channel in fluid communication with a source inlet substantially parallel to the sink channel; and   c) at least one blood vessel chamber disposed between the sink channel and the source channel and comprising at least one first terminal end in fluid communication with the sink channel and at least one second terminal end in fluid communication with the source channel,   wherein the said blood vessel chamber is arranged to correspond to a thickness and length of a blood vessel to be generated, and arranged to form a blood vessel-generating region by filling a gel and/or an extracellular matrix (ECM).   
     
     
         2 . A method of generating a blood vessel comprising:
 a) tilling a gel and/or an ECM into the blood vessel chamber of the blood vessel-generating device according to  claim 1 ;   b) injecting a blood vessel-forming cell into the sink channel and/or the source channel of the blood vessel-generating device, and attaching the blood vessel-forming cell to the gel and/or the ECM exposed to the first terminal end and second terminal end of the blood vessel chamber; and   c) injecting an angiogenesis factor and a cell culture medium into the sink channel and/or the source channel of the blood vessel-generating device, and culturing the blood vessel-forming cell.   
     
     
         3 . The method according to  claim 2 , wherein the gel is at least one selected from the group consisting of a collagen gel, a fibrin gel, Matrigel, a self-assembled peptide gel, a polyethylene glycol gel and an alginate gel. 
     
     
         4 . The method according to  claim 2 , wherein a mixture obtained by mixing the gel and/or the ECM with at least one of the blood vessel-forming cell, the angiogenesis factor, the cell culture medium and the co-culturing cell is filled in step (a). 
     
     
         5 . The method according to  claim 2 , wherein the blood vessel-forming cell is at least one selected from the group consisting of an endothelial cell, an epithelial cell and a cancer cell. 
     
     
         6 . The method according to  claim 2 , further comprising injecting the co-culturing cell into the sink channel and/or the source channel of the blood vessel-generating device, and co-culturing the co-culturing cell. 
     
     
         7 . The method according to  claim 6 , wherein the co-culturing cell is at least one selected from the group consisting of an astrocyte, a glial cell, a mesothelial cell, a fibroblast, a smooth muscle cell and a cancer cell. 
     
     
         8 . A blood vessel-generating device comprising:
 a) a sink channel in fluid communication with a sink inlet;   b) a source channel in fluid communication with a source inlet substantially parallel to the sink channel;   c) a blood vessel-Harming channel in fluid communication with a blood vessel-forming channel inlet, disposed in contact with the sides of the sink channel and the source channel between the sink channel and the source channel, substantially parallel to the source channel and the sink channel;   d) a first culture channel in fluid communication with a first culture channel inlet, disposed in contact with the other side of the sink channel, substantially parallel to the sink channel; and   e) a second culture channel in fluid communication with a second culture channel inlet, disposed in contact with the other side of the source channel, substantially parallel to the source channel,   wherein a plurality of microstructures configured to allow interaction between biochemical materials included in each channel are arranged in the interfaces of the respective adjacent channels at set intervals.   
     
     
         9 . A method of generating a blood vessel comprising:
 a) injecting i) an gel and/or an ECM and ii) a blood vessel-forming cell into the blood vessel-forming channel of the blood vessel-generating device according to  claim 8 ;   b) injecting i) the gel and/or the ECM and ii) a co-culturing cell into the first culture channel and/or the second culture channel of the blood vessel-generating device; and   c) injecting an angiogenesis factor and a cell culture medium into the sink channel and/or the source channel of the blood vessel-generating device, and culturing the blood vessel-forming cell and the co-culturing cell.   
     
     
         10 . A method of generating a blood vessel comprising:
 a) injecting a gel and/or an ECM into the blood vessel-forming channel of the blood vessel-generating device according to  claim 8 ;   b) injecting a blood vessel-forming cell into the sink channel and/or the source channel of the blood vessel-generating device, and attaching the blood vessel-forming cell to the gel and/or the ECM exposed between the microstructures arranged in the sides of the blood vessel-forming, channel;   c) injecting i) the gel and/or the ECM and ii) a co-culturing cell into the first culture channel and/or the second culture channel of the blood vessel-generating device; and   d) injecting an angiogenesis factor and a cell culture medium into the sink channel and/or the source channel of the blood vessel-generating device, and culturing the blood vessel-forming cell and the co-culturing cell.   
     
     
         11 . The method according to  claim 9  or  10 , wherein the blood vessel-forming cell is at least one selected from the group consisting of an endothelial cell, an epithelial cell and a cancer cell. 
     
     
         12 . The method according to  claim 9  or  10 , wherein the gel is at least one selected from the group consisting of a collagen gel, a fibrin gel, Matrigel, a self-assembled peptide gel, a polyethylene glycol gel and an alginate gel. 
     
     
         13 . The method according to  claim 9  or  10 , wherein the co-culturing cell is at least one selected from the group consisting of an astrocyte, a glial cell, a mesothelial cell, a fibroblast, a smooth muscle cell and a cancer cell.