In vitro model for a tumor microenvironment

Abstract

Methods for mimicking a tumor microenvironment in vitro are provided. The methods comprise indirectly applying a shear stress upon at least one tumor cell type plated on a surface within a cell culture container. Methods for mimicking tumor metastasis and methods for testing drugs or compounds in such systems are also provided.

What is claimed is: 
     
         1 . A method for mimicking a tumor microenvironment in vitro, the method comprising:
 adding a culture medium to a cell culture container;   plating at least one tumor cell type on a surface within the cell culture container; and   indirectly applying a shear stress upon the at least one tumor cell type, the shear stress resulting from flow of the culture medium induced by a flow device, the flow mimicking flow to which the tumor cells are indirectly exposed in vivo in the tumor microenvironment, wherein the flow is time-variant.   
     
     
         2 . The method of  claim 1 , further comprising:
 depositing at least one extracellular matrix component on the surface within the cell culture container and plating the at least one tumor cell type on the at least one extracellular matrix component; or   suspending the at least one tumor cell type in a solution comprising at least one extracellular matrix component to create a suspension comprising the at least one tumor cell type and the at least one extracellular matrix component, and depositing the suspension on the surface within the cell culture container; and   indirectly applying the shear stress upon the at least one extracellular matrix component and the at least one tumor cell type.   
     
     
         3 . The method of  claim 1 , further comprising plating the at least one tumor cell type on a first surface of a porous membrane and indirectly applying the shear stress upon the at least one tumor cell type by applying the shear stress upon a second surface of the porous membrane. 
     
     
         4 . A method for mimicking a tumor microenvironment in vitro, the method comprising:
 adding a culture medium to a cell culture container;   plating at least one tumor cell type on a first surface of a porous membrane within the cell culture container; and   indirectly applying a shear stress upon the at least one tumor cell type by applying a shear stress upon a second surface of the porous membrane, the shear stress resulting from flow of the culture medium induced by a flow device, the flow mimicking flow to which the tumor cells are indirectly exposed in vivo in the tumor microenvironment.   
     
     
         5 . A method for mimicking a tumor microenvironment in vitro, the method comprising:
 adding a culture medium to a cell culture container;   plating at least one tumor cell type or stromal cell type on a first surface of a porous membrane within the cell culture container, wherein when the stromal cell type is plated on the first surface of the porous membrane, at least one tumor cell type is present on a surface within the cell culture container; and   indirectly applying a shear stress upon the at least one tumor cell type by applying a shear stress upon a second surface of the porous membrane, the shear stress resulting from flow of the culture medium induced by a flow device, the flow mimicking flow to which the tumor cells are indirectly exposed in vivo in the tumor microenvironment.   
     
     
         6 . The method of  claim 3 , wherein the porous membrane is suspended in the cell culture container such that the first surface is proximal and in spaced relation to a bottom surface of the cell culture container, thereby defining within the cell culture container a lower volume comprising the at least one tumor cell type and an upper volume comprising a second surface of the porous membrane, wherein the shear stress is applied upon the second surface of the porous membrane in the upper volume of the container. 
     
     
         7 . The method of  claim 3 , further comprising
 depositing at least one extracellular matrix component on the first surface of the porous membrane and plating the at least one tumor cell type on the at least one extracellular matrix component; or   suspending the at least one tumor cell type in a solution comprising at least one extracellular matrix component to create a suspension comprising the at least one tumor cell type and the at least one extracellular matrix component, and depositing the suspension on the first surface of the porous membrane;   the porous membrane being suspended in the cell culture container such that the first surface is proximal and in spaced relation to a bottom surface of the cell culture container, thereby defining within the cell culture container a lower volume comprising the at least one extracellular matrix component and the at least one tumor cell type, and an upper volume comprising a second surface of the porous membrane, wherein the shear stress is applied upon the second surface of the porous membrane in the upper volume of the container.   
     
     
         8 . The method of  claim 3 , further comprising plating endothelial cells on the second surface of the porous membrane and applying the shear stress upon the plated endothelial cells, wherein the at least one tumor cell type is plated on a first surface of the porous membrane. 
     
     
         9 . The method of  claim 3 , further comprising plating at least one stromal cell type on the second surface of the porous membrane and applying the shear stress upon the plated stromal cell type. 
     
     
         10 . The method of  claim 9 , further comprising plating endothelial cells on the second surface of the porous membrane. 
     
     
         11 . The method of  claim 10 , comprising mixing the at least one stromal cell type with the endothelial cells prior to plating, and applying the shear stress upon the plated mixture of the at least one stromal cell type and the endothelial cells. 
     
     
         12 . The method of  claim 10 , comprising sequentially plating the at least one stromal cell type and the endothelial cells on the second surface of the porous membrane. 
     
     
         13 . The method of  claim 12 , comprising plating the at least one stromal cell type on the second surface of the porous membrane, subsequently plating the endothelial cells on the plated stromal cell type, and applying the shear stress on the plated endothelial cells. 
     
     
         14 . The method of  claim 12 , comprising plating the endothelial cells on the second surface of the porous membrane, subsequently plating the at least one stromal cell type on the plated endothelial cells, and applying the shear stress on the plated stromal cell type. 
     
     
         15 . The method of  claim 3 , wherein the porous membrane is a first porous membrane and the method comprises plating the at least one tumor cell type on a first surface of the first porous membrane, plating at least one stromal cell type on a second surface of the first porous membrane, placing a second porous membrane on the plated stromal cell type such that a first surface of the second porous membrane contacts the plated stromal cells, and applying the shear force upon a second surface of the second porous membrane. 
     
     
         16 . The method of  claim 7 , wherein the porous membrane is a first porous membrane and the method comprises:
 depositing the at least one extracellular matrix component on the first surface of the first porous membrane and plating the at least one tumor cell type on the at least one extracellular matrix component; or   depositing the suspension comprising the at least one tumor cell type and the at least one extracellular matrix component on the first surface of the first porous membrane; and   plating at least one stromal cell type on a second surface of the first porous membrane, placing a second porous membrane on the plated stromal cell type such that a first surface of the second porous membrane contacts the plated stromal cells, and applying the shear force upon a second surface of the second porous membrane.   
     
     
         17 . The method of  claim 15 , further comprising plating endothelial cells on the second surface of the second porous membrane and applying the shear force upon the plated endothelial cells. 
     
     
         18 . The method of  claim 16 , further comprising plating endothelial cells on the second surface of the second porous membrane and applying the shear force upon the plated endothelial cells. 
     
     
         19 . The method of  claim 5 , wherein the porous membrane is a first porous membrane and the method comprises:
 plating the at least one stromal cell type on a first surface of a first porous membrane;   placing a second porous membrane on the plated stromal cell type, such that a first surface of the second porous membrane contacts the plated stromal cells;   plating at least one tumor cell type on a second surface of the second porous membrane; and   indirectly applying the shear stress upon the at least one tumor cell type by applying the shear stress upon the second surface of the first porous membrane.   
     
     
         20 . The method of  claim 18 , wherein the first porous membrane is suspended in the cell culture container such that the first surface of the first porous membrane is proximal and in spaced relation to a bottom surface of the cell culture container, thereby defining within the cell culture container a lower volume comprising the at least one tumor cell type, the second porous membrane, and the at least one stromal cell type, and an upper volume comprising a second surface of the first porous membrane, wherein the shear stress is applied upon the second surface of the first porous membrane in the upper volume of the container.