Methods, Devices and Systems for Enhanced Transduction Efficiency

Abstract

The systems and methods are directed to leveraging the channel geometry and configuration to overcome diffusion limitations of current transduction systems. The methods may include a method of transducing target cells using a device. The device may include at least one continuous channel. The method may include delivering target cells and viral vectors into a transduction region of the channel through a first inlet/outlet simultaneously and/or consecutively when one or more additional inlets/outlets that are located in a region of the channel downstream of the transduction are closed. After transducing for some incubation time, a flushing solution may be delivered through one or more of the additional inlets/outlets. The method may include collecting transduced cells after the transducing incubation time and the delivering of the flushing solution.

What is claimed: 
     
         1 . A method of transducing target cells using a device, the device including at least one continuous channel, and comprising the following steps
 delivering target cells and viral vectors into a transduction region of the channel through a first inlet/outlet simultaneously and/or consecutively when one or more additional inlets/outlets that are located in a region of the channel downstream of the transduction are closed;   after transducing for some incubation time, delivering a flushing solution through one or more of the additional inlets/outlets; and   collecting transduced cells after the transducing incubation time and the delivering of the flushing solution.   
     
     
         2 . The method according to  claim 1 , further comprising:
 providing the device, the device including:
 the transduction region; 
 a flushing region; and 
 a connection region disposed between the transduction region and the flushing region, 
   wherein the transduction region, the flushing region, and the connection region forms a continuous channel.   
     
     
         3 . The method according to  claim 2 , wherein:
 wherein the first inlet/outlet is disposed at a first end of the channel in the transduction region; and   wherein the one or more additional inlets/outlets are disposed in the transduction and/or connection region.   
     
     
         4 . The method according to  claim 1 , wherein:
 the one or more additional inlets/outlets include a second inlet/outlet and a third outlet that is disposed on the channel further downstream than the second port;   the delivering of the target cells and/or the viral vector through the first inlet/outlet when the third port is outlet;   the delivering of the flushing solution is through the second inlet/outlet and/or the third inlet/outlet; and   excess cells and/or viral vectors can be collected through the second inlet/outlet.   
     
     
         5 . The method according to  claim 4 , wherein the delivering of the flushing solution is through the second inlet/outlet when the third inlet/outlet is closed and the method further comprises:
 closing the second inlet/outlet; and   delivering a flushing solution through the third inlet/outlet,   wherein the collecting of the transduced cells is after the delivering of the flushing solution through the second inlet/outlet and third inlet/outlet.   
     
     
         6 . The method according to  claim 5 , further comprising:
 providing the device, the device including:
 the transduction region; 
 a flushing region; and 
 a connection region disposed between the transduction region and the flushing region. 
   wherein the transduction region, the flushing region, and the connection region forms a continuous channel;   wherein the second inlet/outlet is disposed between the flushing region and the connection region; and   wherein the third inlet/outlet is disposed at the end of the flushing region.   
     
     
         7 . The method according to  claim 6 , wherein:
 wherein the channel disposed in the transduction region is coated with a removable, cell adhesive coating.   
     
     
         8 . The method according to  claim 7 , wherein the coating is RectroNectin. 
     
     
         9 . The method according to  claim 6 , wherein:
 the flushing region includes a split portion in which the channel splits into two segments; and the flushing through the third inlet/outlet causes the fluid to flush out the cells along sides of the channel.   
     
     
         10 . The method according to  claim 6 , wherein the channel has height of about 50-100 μm and a width within about 3000-5000 μm. 
     
     
         11 . A device configured for transducing target cells with viral vector; the device including:
 a transduction region;   a flushing region;   a connection region disposed between the transduction region and the flushing region, wherein the transduction region, the flushing region, and the connection region forms a continuous channel; and   one or more inlets/outlets disposed at end of the transduction region and one or more inlets/outlets disposed on the flushing region;   wherein the channel has a height and a width greater than the height.   
     
     
         12 . The device according to  claim 11 , wherein the channel has height of about 50-100 μm and a width within about 3000-5000 μm. 
     
     
         13 . The device according to  claim 11 , wherein the channel has height of about 50-250 μm and a width of about 1000-5000 μm. 
     
     
         14 . The device according to  claim 11 , wherein channel has a total surface area of about 9.5-15 cm 2 . 
     
     
         15 . The device according to  claim 11 , further comprising:
 a second inlet/outlet disposed between the flushing region and the connection region; and   a third inlet/outlet is disposed at an end of the flushing region;   wherein the second inlet/outlet is disposed between the first inlet/outlet and the third inlet/outlet.   
     
     
         16 . The device according to  claim 15 , wherein:
 the flushing region extends between the second inlet/outlet and the third inlet/outlet; and   the flushing region includes a splitting portion in which the channel splits into two sections.   
     
     
         17 . The device according to  claim 11 , wherein:
 wherein the channel disposed in the transduction region is coated with a removable, cell adhesive coating.   
     
     
         18 . The device according to  claim 17 , wherein the coating is RectroNectin. 
     
     
         19 . The device according to  claim 11 , further comprising:
 one or more adaptors, the one or more adapters configured to communicate with each inlet/outlet.   
     
     
         20 . The device according to  claim 19 , wherein the one or more adapters are disposed on the inlet/outlet and/or on a side of the device.