High throughput cell migration assay plates and methods of fabrication

Abstract

A Cell Migration Assay Plates (CMAP) assembly for high throughput microfluidic migration assays and method of manufacturing thereof are provided. The CMAP assembly includes a top plate having a plurality of wells aligned with a bottom plate having a plurality of troughs. Each of the plurality of wells is defined at least in part by first and second reservoirs and a divisional wall extending between the reservoirs. The bottom plate is secured to the top plate to form a plurality of micro-channels, such that each one of the plurality of micro-channels is defined by a portion of one of the divisional walls and a portion of a corresponding one of the plurality of troughs. The plurality of micro-channels enable communication between the reservoirs and visualization of cells migrating through the micro-channels. In this manner, migration of cells through the micro-channels can be visualized for testing and screening applications.

What is claimed is: 
     
         1 . A cell migration assay plates (CMAP) assembly comprising:
 a top plate having a plurality of wells, each of the plurality of wells defined at least in part by a first reservoir, a second reservoir, and a divisional wall extending between the first reservoir and the second reservoir;   a bottom plate operable to be secured to the top plate, the bottom plate having a plurality of troughs; and   a plurality of micro-channels formed when the bottom plate is secured to the top plate, each of the plurality of micro-channels defined by a portion of the divisional wall of each of the plurality of wells and middle portions of the plurality of troughs, the plurality of micro-channels enabling communication between the first reservoir and the second reservoir when the bottom plate is secured to the top plate.   
     
     
         2 . The CMAP assembly of  claim 1 ,
 wherein,
 each of the middle portions of the plurality of troughs is connected to an entry portion and an exit portion, 
 the entry portion and the exit portion extend away from the divisional wall on opposite sides of the divisional wall, when the bottom plate is secured to the top plate. 
   
     
     
         3 . The CMAP assembly of  claim 1 ,
 wherein,
 the first reservoir and the second reservoir are elongated reservoirs, and 
 the first reservoir extends parallel to the second reservoir. 
   
     
     
         4 . The CMAP assembly of  claim 1 ,
 wherein,
 the plurality of troughs includes a plurality of trough sets, and 
 each of the plurality of trough sets includes a first linear array of troughs and a second linear array of troughs. 
   
     
     
         5 . The CMAP assembly of  claim 4 ,
 wherein,
 each of the plurality of troughs sets extend between a respective one of the plurality of wells. 
   
     
     
         6 . The assembly of  claim 1 ,
 wherein,
 the communication is one-way communication from the first reservoir to the second reservoir, 
 the first reservoir is a seeding reservoir for tumor cells and/or one or more drugs in each of the plurality of wells, and 
 the second reservoir is an output reservoir to which the tumor cells or the one or more drugs are operable to migrate via a three-dimensional (3D) migration without application of any gradient between the first reservoir and the second reservoir in response being physical confined by the plurality of micro-channels. 
   
     
     
         7 . The assembly of  claim 1 , wherein the plurality of wells include four (4) wells or ninety-six (96) wells. 
     
     
         8 . The assembly of  claim 1 , wherein each of the plurality of micro-channels include two-hundred and forty (240) micro-channels. 
     
     
         9 . The assembly of  claim 1 , wherein the plurality of micro-channels have varying lengths. 
     
     
         10 . The assembly of  claim 1 , wherein lengths of the plurality of micro-channels vary from 100 μm to 2.0 mm. 
     
     
         11 . The assembly of  claim 1 , wherein the plurality of micro-channels have varying aspect ratios of height-to-width. 
     
     
         12 . The assembly of  claim 1 , wherein the plurality of micro-channels have dimensions decreasing continuously. 
     
     
         13 . The assembly of  claim 1 , wherein the plurality of micro-channels have dimensions reduced at discrete steps. 
     
     
         14 . The assembly of  claim 1 , wherein the plurality of micro-channels have dimensions increasing continuously. 
     
     
         15 . The assembly of  claim 1 , wherein the plurality of micro-channels have dimensions increased at discrete steps. 
     
     
         16 . The assembly of  claim 1 , wherein the divisional wall has a thickness ranging from 100 μm to 400 μm. 
     
     
         17 . The assembly of  claim 1 , wherein each of the plurality of wells is a microfluidic device and comprises 50 to 400 micro-channels such that the CMAP assembly has a high throughput. 
     
     
         18 . The assembly of  claim 1 , wherein the plurality of micro-channels are sized for tumor cells to squeeze through the micro-channels to provide three-dimensional (3D) migration. 
     
     
         19 . The assembly of  claim 1 , wherein each of the plurality of troughs includes opposing side walls and a bottom wall extending between the opposing side walls. 
     
     
         20 . The assembly of  claim 1 , wherein each of the plurality of micro-channels includes a square cross-section or a rectangular cross-section. 
     
     
         21 . A method of fabricating a cell migration assay plates (CMAP) assembly, the method comprising:
 forming a top plate having a plurality of wells, each of the plurality of wells defined at least in part by a first reservoir, a second reservoir, and a divisional wall extending between the first reservoir and the second reservoir;   forming a bottom plate having a plurality of troughs; and   forming a plurality of micro-channels by securing the top plate to the bottom plate, each of the plurality of micro-channels defined by a portion of the divisional wall of each of the plurality of wells and middle portions of the plurality of troughs, the plurality of micro-channels enabling communication between the first reservoir and the second reservoir when the bottom plate is secured to the top plate.   
     
     
         22 . The method of  claim 21 , further including:
 aligning the top plate and the bottom plate so the divisional wall abuts the plurality of troughs.   
     
     
         23 . The method of  claim 21 ,
 wherein,
 the top plate is formed via an injection molding process; and 
 the bottom plate is formed via a hot embossing process. 
   
     
     
         24 . The method of  claim 21 , wherein the bottom plate is formed using an optically translucent or transparent biocompatible cyclic olefin polymer (COP) to enable viewing of cell migration in the micro-channels. 
     
     
         25 . The method of  claim 21 , wherein the top plate is formed using a black cyclic olefin polymer (COP).