Multicellular system and method for multiparametric analysis

Abstract

A system for multiparametric analysis includes a substrate and a three-dimensional (3D) cell aggregate. The substrate has a major surface and includes at least one radial electrode array. The 3D cell aggregate is disposed on the major surface of the substrate. The 3D cell aggregate has a longitudinal surface at least a portion of which covers one or more of the electrodes of the radial electrode array.

1 - 7 . (canceled) 
     
     
         8 . A method comprising the steps of:
 providing a multiparametric analysis system comprising a substrate and a 3D cell aggregate, the substrate having a major surface and including at least one radial electrode array, the 3D cell aggregate being disposed on the major surface of the substrate and having a longitudinal surface at least a portion of which covers one or more of the electrodes comprising the radial electrode array;   contacting at least one test agent with the 3D cell aggregate; and   measuring one or more responses of the 3D cell aggregate to the at least one test agent.   
     
     
         9 . The method of  claim 8 , wherein the measuring step further includes determining the level of one or more diffusing analytes. 
     
     
         10 . The method of  claim 9 , wherein the diffusing analyte includes oxygen or hydrogen ions. 
     
     
         11 . The method of  claim 8 , wherein the measuring step includes determining the level of one or more biomarkers associated with cellular metabolism. 
     
     
         12 . The method of  claim 11 , wherein the biomarker is a hypoxia-related marker and/or a cancer biomarker. 
     
     
         13 . The method of  claim 8 , wherein the at least one test agent is a biological and/or chemical compound. 
     
     
         14 . The method of  claim 10 , wherein a continuous model is used to determine oxygen concentration within the 3D cell aggregate whereby the oxygen consumption-concentration relationship is linearly proportional. 
     
     
         15 . The method of  claim 8 , wherein one or more of the measured responses is used to identify a potential therapeutic agent. 
     
     
         16 - 17 . (canceled) 
     
     
         18 . The method of  claim 8 , wherein the substrate has a multilayer configuration comprising:
 a base layer;   an intermediate layer disposed on at least a portion of the base layer including the radial electrode array, wherein the radial electrode array comprises spiral electrodes; and   a spacer layer comprising the major surface disposed on at least a portion of the base layer to mitigate fouling of the spiral electrodes.   
     
     
         19 . The method of  claim 18 , wherein the spiral electrodes comprise Fermat's geometry 
     
     
         20 . The method of  claim 18 , wherein the spiral electrodes obtain information used to detect one or more biomarkers in the 3D cell aggregate. 
     
     
         21 . The method of  claim 8 , wherein the 3D cell aggregate has a microliter volume. 
     
     
         22 . The method of  claim 21 , wherein the 3D cell aggregate has a longitudinal surface at least a portion of which covers one or more of the electrodes. 
     
     
         23 . The method of  claim 22 , wherein the 3D cell aggregate is heterogeneous comprising at least two cell types. 
     
     
         24 . The method of  claim 23 , wherein the cell types comprise a non-cancerous cell type and a cancerous cell type. 
     
     
         25 . The method of  claim 8 , the measuring step further includes determining the one or more responses of the 3D cell aggregate to the at least one test agent based on one or more of oxygen tension, hydrogen ion concentration, and drug concentration within the 3D cell aggregate. 
     
     
         26 . The method of  claim 8 , wherein the substrate is prepared using a MEMS microfabrication technique. 
     
     
         27 . The method of  claim 8 , wherein the 3D cell aggregate represents an in vivo environment comprising a portion of tissue. 
     
     
         28 . The method of  claim 8 , wherein the array comprises a diametric array of spiral electrodes along a diameter of the 3D cell aggregate.