Electrophysiological recording system and methods of using same

Patent metadata

Field	Value
Publication number	US-9448195-B2
Application number	US-201113810402-A
Country	US
Kind code	B2
Filing date	Jul 15, 2011
Priority date	Jul 16, 2010
Publication date	Sep 20, 2016
Grant date	Sep 20, 2016

Abstract

Electrophysiological recording systems for analysis of cultured cells include an recording device and a housing for supporting the electrophysiological recording device. The device has a porous membrane with top and bottom surfaces and a plurality of pores extending between the top and bottom surfaces. The porous membrane defines a first cell culture region disposed on the top surface and an opposed second cell culture region disposed on the bottom surface. The electrophysiological recording device has a plurality of electrodes positioned on the porous membrane in between a first insulation layer and a second insulation layer. The plurality of electrodes extend to the first cell culture region of the porous membrane. Recording ends of the electrodes measure electrical properties of cells cultured within the first and second cell culture regions, while the contact ends of the electrodes are positioned in electrical communication with data acquisition equipment.

First claim

What is claimed is:

1. An electrophysiological recording system for analysis of cultured cells, the electrophysiological recording system comprising:
an electrophysiological recording device comprising:
a porous membrane having a top surface and an opposed bottom surface, the porous membrane defining a first cell culture region disposed on a select portion of the top surface and an opposed second cell culture region disposed on a select portion of the bottom surface, the porous membrane further defining a plurality of pores extending between the opposed first and second cell culture regions on the respective opposed top and bottom surfaces; and
an 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 top and bottom surfaces, wherein the contact ends of the plurality of electrodes are configured for electrical communication with data acquisition equipment, and 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; and

a housing defining a cell culture chamber, wherein at least a portion of the recording portion of the porous membrane is mountable within the cell culture chamber of the housing,
wherein the porous membrane and the electrode array of the electrophysiological recording device cooperate to form a cell culture structure and are configured to be rolled up together from a substantially planar configuration to form a three-dimensional cell culture structure having a spiral cross-sectional shape, and wherein the housing comprises means for rolling up the porous membrane and the electrode array to form the three-dimensional cell culture structure.

2. The electrophysiological recording system of claim 1 , further comprising means for culturing cells on the top surface and the bottom surface of the porous membrane within the opposed first and second cell culture regions.

3. The electrophysiological recording system of claim 2 , wherein the cells cultured on the top surface of the porous membrane differ from the cells cultured on the bottom surface of the porous membrane.

4. The electrophysiological recording system of claim 1 , further comprising a Petri dish, wherein the cell culture chamber of the housing is configured for receipt therein the Petri dish.

5. The electrophysiological recording system of claim 1 , wherein the respective contact ends of adjacent electrodes of the plurality of electrodes are spaced apart from one another at at least one predetermined distance and the respective recording ends of adjacent electrodes of the plurality of electrodes are spaced apart from one another at at least one predetermined distance.

6. The electrophysiological recording system of claim 1 , wherein the porous membrane has a thickness ranging from about 5 micrometers to about 15 micrometers.

7. The electrophysiological recording system of claim 1 , wherein each pore of the plurality of pores of the porous membrane has a diameter ranging from about 2 micrometers to about 4 micrometers.

8. The electrophysiological recording system of claim 1 , wherein the plurality of pores of the porous membrane are present at a density ranging from about 150,000 pores per square centimeter to about 250,000 pores per square centimeter.

9. The electrophysiological recording system of claim 8 , wherein the plurality of pores of the porous membrane are randomly scattered within the opposed first and second cell culture regions.

10. The electrophysiological recording system of claim 1 , further comprising means for inducing an action potential in cells cultured within at least one of the first cell culture region and the second cell culture region.

11. The electrophysiological recording system of claim 1 , wherein the means for inducing an action potential comprises at least one electrode of the plurality of electrodes that is configured to stimulate electrical activity within the recording portion of the porous membrane.

12. The electrophysiological recording system of claim 1 , wherein the means for rolling up the porous membrane and the electrode array is configured to form a three-dimensional cell culture having a substantially spiral cross-sectional shape.

13. The electrophysiological recording system of claim 1 , wherein the means for rolling up the porous membrane and the electrode array comprises a gear assembly that comprises: a rod configured for attachment 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.

14. The electrophysiological recording system of claim 13 , wherein the rod comprises a rod electrode configured for measurement of electrical properties of the formed three-dimensional cell culture and for electrical communication with the data acquisition equipment.

15. The electrophysiological recording system of claim 1 , wherein the electrophysiological recording device further comprises:
a first insulation layer positioned therebetween at least a portion of each electrode of the plurality of electrodes and the top surface of the porous membrane; and
a second insulation layer positioned thereon at least a portion of each electrode of the plurality of electrodes such that a lead portion of each electrode is positioned therebetween the first insulation layer and the second insulation layer.

16. The electrophysiological recording system of claim 15 , wherein the first insulation layer is applied thereon the top surface of the porous membrane and thereon at least a portion of the plurality of pores defined by the porous membrane.

17. The electrophysiological recording system of claim 16 , wherein the first insulation layer is applied as a continuous layer across a portion of the top surface of the porous membrane and within and across at least a portion of the plurality of pores defined by the porous membrane.

18. The electrophysiological recording system of claim 15 , wherein at least one of the first insulation layer and the second insulation layer comprises parylene.

19. The electrophysiological recording system of claim 1 , wherein the housing comprises:
a first base support portion defining an opening;
a second base support portion configured to support 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 configured for attachment thereto 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.

20. The electrophysiological recording syst

Assignees

• Moreno Alonso P
• Goldberg Gary S
• Mondal Abhijit
• Harvey Ian
• Baker Brian
• Univ Utah Res Found

Inventors

• Moreno Alonso P
• Goldberg Gary S
• Mondal Abhijit
• Harvey Ian
• Baker Brian

Classifications

• C12N2535/00

  Supports or coatings for cell culture characterised by topography · CPC title

• C12N5/0068 Primary

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• C12N2537/00

  Supports and/or coatings for cell culture characterised by physical or chemical treatment · CPC title

• G01N27/00 Primary

  Investigating or analysing materials by the use of electric, electrochemical, or magnetic means (G01N3/00 – G01N25/00 take precedence; measurement or testing of electric or magnetic variables or of electric or magnetic properties of materials G01R) · CPC title

• C12M25/02 Primary

  Membranes; Filters (filters or filtration in general B01D24/00-B01D41/00) · CPC title