Electrophysiological recording system and methods of using same

Abstract

The disclosure provided herein provides systems, devices, and methods for analyzing cultured cells. Also disclosed herein are systems, devices, and methods for analyzing interactions between two different groups of cells. Also disclosed herein are systems and methods of producing and analyzing a three-dimensional cell culture.

What is claimed is: 
     
         1 . A method of forming a three-dimensional cell culture, comprising:
 positioning a porous membrane having a first surface and an opposed second surface in a substantially planar configuration, the porous membrane defining a first cell culture region disposed on a select portion of the first surface and a second cell culture region disposed on a select portion of the second surface, the porous membrane defining a plurality of pores extending between the first surface and the second surface within the opposed first and second cell culture regions, wherein an electrode array is secured to the porous membrane, the electrode array comprising a plurality of electrodes having distal recording ends that are secured to the porous membrane and extend to the first cell culture region of the porous membrane, and wherein the porous membrane and the electrode array cooperate to form a cell culture structure;   applying a first group of cells thereon the first surface of the porous membrane within the first cell culture region;   applying a second group of cells thereon the opposed second surface of the porous membrane within the second cell culture region; and   rolling up the porous membrane and the electrode array to form a three-dimensional cell culture structure having a spiral cross-sectional shape.   
     
     
         2 . The method of  claim 1 , further comprising applying one or more physiological substrates to at least one of the first surface or the second surface of the porous membrane prior to applying the first group of cells and the second group of cells. 
     
     
         3 . The method of  claim 1 , wherein the first group of cells differs from the second group of cells, wherein the first group of cells comprises a first cell type and the second group of cells comprises a second cell type different than the first cell type. 
     
     
         4 . The method of  claim 3 , wherein the first cell type comprises cardiac myocytes. 
     
     
         5 . The method of  claim 4 , wherein the second cell type comprises at least one of myofibroblasts or fibroblasts. 
     
     
         6 . The method of  claim 1 , wherein the first group of cells and the second group of cells comprise one or more of neurons, oligodendrocytes, and astrocytes. 
     
     
         7 . The method of  claim 1 , wherein the first group of cells comprises smooth muscle cells. 
     
     
         8 . The method of  claim 7 , wherein the second group of cells comprises endothelial cells. 
     
     
         9 . The method of  claim 1 , further comprising positioning the porous membrane in a cell culture chamber. 
     
     
         10 . The method of  claim 9 , further comprising securing the porous membrane to a housing, wherein the housing comprises:
 a first base support portion defining an opening;   a second base support portion that supports the porous membrane such that at least a portion of the opposed first and second cell culture regions of the porous membrane overlie the opening of the first base support portion; and   a first cover portion defining an opening, the first cover portion being attached to the first base support portion such that the opening of the first cover portion is substantially aligned with the opening of the first base support portion,   wherein the respective openings of the first base support portion and the first cover portion cooperate to define the cell culture chamber of the housing.   
     
     
         11 . The method of  claim 10 , further comprising positioning at least the first base support portion of the housing within a container holding the cell culture chamber. 
     
     
         12 . The method of  claim 11 , wherein the container is a Petri dish. 
     
     
         13 . The method of  claim 1 , further comprising detecting at least one electrical signal at at least one recording end of the plurality of electrodes of the electrode array indicative of one or more electrical properties of the first and second regions of cells. 
     
     
         14 . The method of  claim 13 , further comprising positioning the porous membrane in a cell culture chamber such that the at least one recording end of the plurality of electrodes contacts the cell culture chamber. 
     
     
         15 . The method of  claim 13 , further comprising transmitting the at least one electrical signal to data acquisition equipment. 
     
     
         16 . The method of  claim 15 , wherein the at least one electrical signal is transmitted to the data acquisition equipment prior to formation of the three-dimensional cell culture structure. 
     
     
         17 . The method of  claim 15 , wherein the at least one electrical signal is transmitted to the data acquisition equipment following formation of the three-dimensional cell culture structure. 
     
     
         18 . The method of  claim 1 , further comprising inducing an action potential in cells cultured within at least one of the first cell culture region and the second cell culture region. 
     
     
         19 . The method of  claim 18 , wherein the step of inducing an action potential comprises activating a light source configured to trigger the action potential. 
     
     
         20 . The method of  claim 19 , wherein the at least one of the first group of cells and the second group of cells comprises cells having light-gated ion channels. 
     
     
         21 . The method of  claim 20 , wherein the cells having light-gated ion channels comprise cells that are transfected with ChannelRhodopsin. 
     
     
         22 . The method of  claim 21 , wherein the first group of cells comprises myocytes, and wherein the second group of cells comprises Hela cells expressing Cx43 and transfected with ChannelRhodopsin-2. 
     
     
         23 . The method of  claim 1 , wherein portions of the first and second cell culture regions proximate the recording ends of the plurality of electrodes define a recording portion of the porous membrane. 
     
     
         24 . The method of  claim 23 , further comprising inducing an action potential using at least one electrode of the plurality of electrodes, wherein the at least one electrode of the plurality of electrodes is configured to stimulate electrical activity within the recording portion of the porous membrane. 
     
     
         25 . The method of  claim 1 , wherein the porous membrane and the electrode array are rolled up using a gear assembly comprising:
 a rod attached to at least a portion of the recording portion of the porous membrane; and   at least one gear coupled to the rod and rotatable about a rotation axis, wherein rotation of the at least one gear about the rotation axis results in a corresponding rotation of the rod.   
     
     
         26 . The method of  claim 25 , further comprising attaching at least a portion of the recording portion of the porous membrane to the rod of the gear assembly. 
     
     
         27 . The method of  claim 26 , further comprising rotating the at least one gear of the gear assembly relative to the rotation axis to effect rotation of the rod and rolling up of the porous membrane. 
     
     
         28 . The method of  claim 26 , wherein the rod comprises an electrode, wherein the porous membrane defines a central aperture that is substantially aligned with a tip of the electrode of the rod, and wherein the method further comprises using the central aperture to diffuse electrolytes within the three-dimensional cell culture. 
     
     
         29 . A method of forming a three-dimensional cell culture, comprising:
 applying a first group of cells thereon a first surface of a porous membrane positioned in a substantially planar configuration, the porous membrane having a second surface opposed from the first surface, the porous membrane defining a first cell culture region disposed on a select portion of the first surface and an opposed second cell culture region disposed on a select portion of the second surface, the porous membrane further defining a plurality of pores extending between the opposed first and second cell culture regions on the respective opposed first and second surfaces, wherein the porous membrane is configured to cooperate with an electrode array to form an electrophysiological recording device, the electrode array comprising a patterned layer that defines a plurality of electrodes, each electrode of the plurality of electrodes having a distal recording end and a proximal contact end, wherein the distal recording end of each electrode of the plurality of electrodes is secured to the porous membrane and extends to the first cell culture region of the porous membrane, wherein the recording ends of the plurality of electrodes are configured for measurement of electrical properties of cells cultured within the first and second cell culture regions on the respective opposed first and second surfaces, wherein the contact ends of the plurality of electrodes are configured for electrical communication with data acquisition equipment;   applying a second group of cells thereon the opposed second surface of the porous membrane; and   rolling up the porous membrane and the electrode array of the electrophysiological recording device to form a three-dimensional cell culture structure having a spiral cross-sectional shape.