Method and related systems for use with a fluidics device

Abstract

Disclosed are fluidics devices and assemblies allowing for fluid flow between a plurality of wells. The fluidics devices and assemblies that are provided mimic in vivo tissue environments by allowing for initially segregated tissue cultures that can then be linked through fluid flow to measure integrated tissue response. The fluidics devices and assemblies provide a pumpless system using surface tension, gravity, and channel geometries. By linking human tissue functional systems to better simulate in vivo feedback and response signals between the tissues, the need for testing in animals can be minimized.

The invention claimed is: 
     
         1 . An in vitro assay method for determining whether a parameter was diminished or enhanced by a cell culture response to a test compound, comprising:
 applying a dynamic fluid including the test compound to a dynamic well of a fluidics device, wherein the fluidics device further includes a dynamic fluid pathway extending from the dynamic well, and further includes a cell culture positioned in the fluid pathway and/or the dynamic well;   applying a control fluid not including the test compound to a control well of the fluidics device, wherein the fluidics device further includes a control fluid pathway extending from the control well;   performing a dynamic bioassay to measure the parameter of the dynamic well and/or the dynamic fluid pathway and determining a dynamic value;   performing a control bioassay to measure the parameter of the control well and/or the control fluid pathway and determining a control value;   comparing the dynamic value to the control value for determining whether the parameter was diminished or enhanced by the cell culture response to the test compound or a metabolite of the test compound.   
     
     
         2 . The method of  claim 1 , further comprising:
 wherein the dynamic value includes a dynamic effective concentration at a percentage X (ECx) for the parameter, wherein 0≦X≦100;   wherein the control value includes a control ECx for the parameter; and   wherein a lower value of the at least one dynamic ECx relative to the control ECx is predictive of the cell culture response enhancing the parameter and a higher value of the at least one dynamic ECx relative to the control ECx is predictive of the cell culture response diminishing the parameter.   
     
     
         3 . The method of  claim 2 , wherein an effect detection reagent is present in the dynamic well or the dynamic fluid pathway having a cell culture therein, and wherein determination of the dynamic ECx includes detection of the effect detection reagent. 
     
     
         4 . The method of  claim 1 , further comprising:
 removing a portion of the dynamic fluid, or a portion thereof, from at least one stop well of the dynamic fluid pathway; and   applying a stop to the at least one stop well for controlling the flow of the dynamic fluid in preparation for the dynamic bioassay.   
     
     
         5 . The method of  claim 1 , wherein the performing of any one or more of the bioassays includes removing an aliquot of the respective fluid from the respective fluid pathway or well at one or more time periods. 
     
     
         6 . The method of  claim 1 , wherein the applying of the dynamic fluid and/or the control fluid is repeated using a specific volume at determined intervals over a specified period of time. 
     
     
         7 . The method of  claim 6 , wherein the applying of the dynamic fluid and/or the control fluid is performed automatically by a robotic liquid handling apparatus or by using a piston assembly nestably engaged with the fluidics device. 
     
     
         8 . The method of  claim 6 , further comprising syphoning the respective fluid from a distal portion of the dynamic fluid pathway and/or the control fluid when the applying of the dynamic fluid and/or the control fluid is repeated. 
     
     
         9 . The method of  claim 8 , where the syphoning the respective fluid is performed automatically by a robotic liquid handling apparatus. 
     
     
         10 . The method of  claim 1 , further comprising:
 applying a tracing fluid having a known concentration of a detectable tracing compound to a tracing well of the fluidics device, wherein the fluidics device further includes a tracing fluid pathway extending from the tracing well, and further includes a cell culture positioned in the tracing fluid pathway and/or the tracing well;   determining the concentration of the tracing compound as a standard curve to calculate a concentration of the test compound or the metabolite of the test compound.   
     
     
         11 . The method of  claim 1 , comprising:
 applying a static fluid including the test compound to a static well;   performing a static bioassay to measure the parameter of the static well and determining a static value;   comparing the dynamic value to the static value for determining whether the parameter was diminished or enhanced by the cell culture response.   
     
     
         12 . The method of  claim 11 , further comprising:
 wherein the dynamic value includes a dynamic effective concentration at a percentage X (ECx) for the parameter, wherein 0≦X≦100;   wherein the static value includes a static ECx for the parameter; and   wherein a lower value of the at least one dynamic ECx relative to the static ECx is predictive of the cell culture response enhancing the parameter and a higher value of the at least one dynamic ECx relative to the static ECx is predictive of the cell culture response diminishing the parameter.   
     
     
         13 . The method of  claim 12 , wherein an effect detection reagent is present in the dynamic well or the dynamic fluid pathway having a cell culture therein, and wherein determination of the dynamic ECx includes detection of the effect detection reagent. 
     
     
         14 . The method of  claim 11 , further comprising:
 removing a portion of the dynamic fluid from at least one stop well of the dynamic fluid pathway; and   applying a stop to the at least one stop well for controlling the flow of the dynamic fluid in preparation for the dynamic bioassay.   
     
     
         15 . The method of  claim 11 , wherein the performing of any one or more of the bioassays includes removing an aliquot of the respective fluid from the respective fluid pathway or well at one or more time periods. 
     
     
         16 . The method of  claim 11 , wherein the applying of the dynamic fluid and/or the control fluid is repeated using a specific volume at determined intervals over a specified period of time. 
     
     
         17 . The method of  claim 16 , wherein the applying of the dynamic fluid and/or the control fluid is performed automatically by a robotic liquid handling apparatus or by using a piston assembly nestably engaged with the fluidics device. 
     
     
         18 . The method of  claim 16 , further comprising syphoning the respective fluid from a distal portion of the dynamic fluid pathway and/or the control fluid when the applying of the dynamic fluid and/or the control fluid is repeated. 
     
     
         19 . The method of  claim 18 , where the syphoning the respective fluid is performed automatically by a robotic liquid handling apparatus. 
     
     
         20 . The method of  claim 11 , further comprising:
 applying a tracing fluid having a known concentration of a detectable tracing compound to a tracing well of the fluidics device, wherein the fluidics device further includes a tracing fluid pathway extending from the tracing well, and further includes a cell culture positioned in the tracing fluid pathway and/or the tracing well;   determining the concentration of the tracing compound as a standard curve to calculate a concentration of the test compound or the metabolite of the test compound.