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Formation and use of neuronal network, and neuron seeding device UPDATE_EN

Foreign code F170009187
File No. AF19-04WO
Posted date Sep 12, 2017
Country United States of America
Application number 201314429035
Gazette No. 20150233890
Gazette No. 9829477
Date of filing Mar 21, 2013
Gazette Date Aug 20, 2015
Gazette Date Nov 28, 2017
International application number JP2013057976
International publication number WO2014045618
Date of international filing Mar 21, 2013
Date of international publication Mar 27, 2014
Priority data
  • P2012-205561 (Sep 19, 2012) JP
  • 2013WO-JP57976 (Mar 21, 2013) WO
Title Formation and use of neuronal network, and neuron seeding device UPDATE_EN
Abstract (US9829477)
Provided are a culture device for neuronal network formation that makes it possible to construct a neuronal network while restricting movement of neurons grown in the culture medium, and a means for the use thereof.
A culture device for neuronal network formation in which is formed a cell fixation part surrounded by a plurality of projections on a flat substrate that can be filled with cell culture medium, and in which (1) there are established spaces having widths such that the neuron cells cannot pass through between the plurality of projections, (2) the inner diameter is such that the cell fixation part can house the cells of from one to a number of neurons, (3) the substrate surface, which is the bottom of the cell fixation part, is coated by an extracellular matrix-forming substance and/or provided with minute through-holes for suctioning culture medium by a suction device on the lower side of the substrate surface.
Scope of claims [claim1]
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.
[claim2]
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.
[claim3]
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.
[claim4]
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.
[claim5]
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.
[claim6]
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.
  • Inventor, and Inventor/Applicant
  • URISU TSUNEO
  • WANG ZHI-HONG
  • UNO HIDETAKA
  • SAITOH MIHO
  • NAGAOKA YASUTAKA
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
IPC(International Patent Classification)
Reference ( R and D project ) CREST Creation of Nanosystems with Novel Functions through Process Integration AREA
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