Formation and use of neuronal network, and neuron seeding device

Patent metadata

Field	Value
Publication number	US-9829477-B2
Application number	US-201314429035-A
Country	US
Kind code	B2
Filing date	Mar 21, 2013
Priority date	Sep 19, 2012
Publication date	Nov 28, 2017
Grant date	Nov 28, 2017

Abstract

A planar patch clamp device is disclosed, which can be used for culturing a neuron in the device so as to form a neuron network, and detecting an electrical property of the neuron that forms the neuron network. The planar patch clamp device includes a plurality of protrusions formed on a first surface, an extracellular matrix forming substance which is coated on the peripheries of a through hole, and electrode sections.

First claim

What is claimed is:

1. A planar patch clamp device for analyzing a neuron network, comprising:
an electrical insulating plate having a first surface, on which a neuron network is formed, a second surface, which is the opposite side, and a through-hole having a diameter which allows passing through both sides of the plate surface, but does not allow the passage of a cell body;
a first liquid pool section formed on the first surface, which enables holding a conductive solution;
a second liquid pool section formed on the second surface, which enables holding a pipet solution, and is communicated with the first liquid pool through the through-hole;
a first surface side-electrode disposed to be electrically conductive to the conductive solution; and
a second surface side-electrode disposed to be electrically conductive to the pipet solution;
wherein a cell plating section surrounded by a plurality of protrusions is formed on the first surface so that the protrusions restrict the movement of cells, but allow the formation of a neuron network, the through-hole is arranged in the cell plating section, and the peripheries of the through-hole are coated with an extracellular matrix-forming substance, and
wherein each of the electrodes on the first surface side and second surface side comprise:
(a) an electrode receptacle for holding each of the electrodes, wherein at least a portion of the receptacle wall that is in contact with the conductive solution or the pipet solution, when the conductive solution or the pipet solution is introduced into the liquid pool sections, is composed of an inorganic porous material;
(b) each of the electrodes having a precious metal chloride (NmCl) layer formed on a surface layer section of the precious metal (Nm), and housed in the electrode receptacle; and
(c) an electrode solution filled into the electrode receptacle, wherein the precious metal chloride (NmCl) and an alkali metal chloride are dissolved at saturated concentration; and
wherein when cells are plated on the cell plating section, both of the first and second surface-side electrodes are not in contact with a cell body.

2. The planar patch clamp device for analyzing a neuron network according to claim 1 , wherein the diameter of the cell plating section defined by the plurality of protrusions is a size that enables accommodating one to several cell bodies.

3. The planar patch clamp device for analyzing a neuron network according to claim 1 , wherein the plurality of cell plating sections defined by the plurality of protrusions are arranged on the plate.

4. The planar patch clamp device for analyzing a neuron network according to claim 1 , one to several cell bodies are placed in the cell placing section, and another cell body is placed outside of the cell placing section.

5. The planar patch clamp device for analyzing a neuron network according to claim 1 , characterized in that the device is used for measurement and analysis of neuronal ion channel current or for imaging analysis.

6. The planar patch clamp device for analyzing a neuron network according to claim 1 further comprising:
a photodetector for detecting light emitted by neurons; and
an irradiating device that irradiates light onto the neurons or plate surface.

Assignees

• Japan Science & Tech Agency

Inventors

• Urisu Tsuneo
• Wang Zhi-Hong
• Uno Hidetaka
• Saitoh Miho
• Nagaoka Yasutaka

Classifications

• G01N33/48728 Primary

  Investigating individual cells, e.g. by patch clamp, voltage clamp (investigating individual particles in general G01N15/10) · CPC title

• C12M23/12

  Well or multiwell plates (C12M25/04 takes precedence) · CPC title

• C12M33/00

  Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus (chemical or physical laboratory apparatus in general B01L, devices for taking cell samples A61B10/0045, withdrawing or distributing predetermined quantities of fluid B01L99/00) · CPC title

• C12N5/0619

  Neurons · CPC title

• C12M23/38

  Caps; Covers; Plugs; Pouring means · CPC title