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Formation and use of neuronal network, and neuron seeding device

Foreign code F170009187
File No. AF19-04WO
Posted date Sep 12, 2017
Country United States of America
Application number 201314429035
Gazette No. 20150233890
Date of filing Mar 21, 2013
Gazette Date Aug 20, 2015
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
Abstract (US20150233890)
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]

1-13. (canceled)
[claim2]
14. 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 cell culture medium;
a second liquid pool section formed on the second surface, which enables holding cell culture medium, and is communicated with the first liquid pool through the through-hole;
a first surface side-electrode disposed to be electrically conductive to the cell culture medium; and
a second surface side-electrode disposed to be electrically conductive to the conducting liquid;
wherein 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 is coated with an extracellular matrix-forming substance.
[claim3]
15. The planar patch clamp device for analyzing a neuron network according to claim 14, wherein the diameter of the cell plating section defined by the plurality of protrusions is a size enables to accommodate one to several cell bodies.
[claim4]
16. The planar patch clamp device for analyzing a neuron network according to claim 14, wherein the plurality of cell plating sections defined by the plurality of protrusions are arranged on the plate.
[claim5]
17. The planar patch clamp device for analyzing a neuron network according to claim 14, one to several cell bodies are placed in the cell placing section, and another cell body is placed outside of the cell placing section.
[claim6]
18. The planar patch clamp device for analyzing a neuron network according to claim 14, characterized in that the electrode sections on the first surface side and second surface side comprises: an electrode receptacle, wherein at least a portion of the receptacle wall that is in contact with the conducting liquid, when the conducting liquid is introduced into the liquid pool sections, is composed of an inorganic porous material;
an electrode 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
an electrode solution filled into the electrode receptacle, wherein the precious metal chloride (NmCl) and an alkali metal chloride are dissolved at saturated concentration.
[claim7]
19. The planar patch clamp device for analyzing a neuron network according to claim 14, characterized in that the device is used for measurement and analysis of neuronal ion channel current or for imaging analysis.
[claim8]
20. The planar patch clamp device for analyzing a neuron network according to claim 14 further comprising: a photodetector for detecting light emitted by neurons; and
an irradiating device that irradiates light onto the neurons or plate surface.
[claim9]
21. A cell seeding device for seeding cell bodies in multiple cell plating sections surrounded by a plurality of protrusions on a plate of a planar patch clamp device, comprising: (1) a suspension supply port for externally supplying cell suspension;
(2) a plurality of fine suspension flow channels extending in a branched manner from the suspension supply port, inside the device body; and
(3) a suspension injection port for injecting the cell suspension into each cell plating section, opened at the bottom of the device body at the end of each suspension flow channel; wherein the neuron seeding device is a board-shaped device body that can be set on the apparatus plate has a size covering the multiple cell plating sections and the flat bottom of the device body is in contact with the apexes of the plurality of protrusions at the multiple cell plating sections in the set situation.
[claim10]
22. The neuron seeding device according to claim 21, characterized in that the plurality of suspension flow channels are set to have substantially the same lengths, and the design is such that when the neuron seeding device is placed on the apparatus plate of the culturing device for formation of a neuron network or the planar patch-clamp device, the individual suspension injection ports are located to precisely correspond with the individual cell plating sections.
[claim11]
23. The neuron seeding device according to claim 21, characterized in that the board-shaped device body comprises an upper board provided with the suspension supply ports and a lower board provided with the suspension injection ports, joined in a closely bonded state, and grooves composing the suspension flow channels are formed on at least one of the bonding surfaces of the upper board and lower board.
[claim12]
24. The neuron seeding device according to claim 21, wherein the device body further comprises a second suspension flow channel system for injection of the neuron suspension into regions other than the cell plating sections of the apparatus plate of the culturing device for formation of a neuron network or the planar patch-clamp device.
  • 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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