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Formation and use of neuronal network, and neuron seeding device

Foreign code F170009194
File No. AF19-04WO
Posted date Sep 12, 2017
Country Republic of Korea
Application number 20157006609
Gazette No. 20150056551
Date of filing Mar 21, 2013
Gazette Date May 26, 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 (KR20150056551)
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.
(From EP2899262 A1)
Scope of claims [claim1]
1. The cell badge to on the charging possible even board be is surrounded with the lug of multiple, forms this cell fixation has the (1)-(3) condition which adds the cell fixation department, the nerve cell network formation cultivation equipment the fact that with feature.
(1) cell fixation department as for under composing between the lug of multiple the cell body of the nerve cell at the time of passage height, do in is and the extended interval is set.
By the lug of (2) multiple inside diameters of the cell fixation department which is provided 1- will be able to accommodate the cell body of nerve cell are the size.
The bottom of (3) cell fixation department the low of composing to the board side had at least one elements in (a) and (b) below.
Cell external matrix formation material (a) coating.
As the badge suction suspicion minute penetration hole by the suction equipment which installed to lower part of (b) minute description board sides, the nerve cell at the time of passage forms the thing of the hole diameter height.
[claim2]
2. There is in a 1st claim,
The minute description cultivation equipment in below (1)-(3) corresponds to which one the nerve cell network formation cultivation equipment the fact that with feature.
As selective territory cell fixation addition is formed as on (1) minute description borads, and as the selective cell nerve cells 1- are arranged as this cell fixation department with and the together different nerve cell only sows to on the borad.
To on (2) minute description borads is formed in cell fixation additional suitable mutual interval of multiple, and to these cell fixation department nerve cells are arranged, 1- with together among the rest as the selective territory which arranged 1 cell fixation additional nerve cell they are used.
Unit of the nerve cell network which corresponds to minute description (1) or (2) to on (3) minute description borads multiple to majority will be able to form, to disperse a minute description cell fixation department to the suitable location from, at the time of formation the high throughput interpretation cultivation equipment of the Keane nerve cell network.
[claim3]
3. The 1st claim or there is in a 2nd claim,
To be a planer patch clamp equipment where the minute description nerve cell network formation cultivation equipment does the nerve cell network in target, and
(1) minute description borads to be the electric dielectric borad, and the bottom of cell fixation department of that electric dielectric board as for under composing the surface of both sides of the board side the chimney height forms the minute description minute penetration hole, the defiant liquid which is a minute description cell badge the nerve cell network formation side which is a first ostensible side of (2) minute penetration holes and, to the second ostensible side which is that opposite side respectively maintains about the defiant liquid of the fluidal undercurrent department for and that fluidal undercurrent department electrification possibly, forms the electrode department which is arranged, the nerve cell suspicion liquid which becomes fixed to fluidal undercurrent addition minute description cell fixation department of (3) first ostensible sides Is becoming with undercurrent department the nerve cell network formation cultivation equipment the fact that with feature.
[claim4]
4. There is in a 3rd claim,
Is in the minute description planer patch clamp equipment, minute description first ostensible side and (a)-(c) element below electrode adding of second ostensible side as for under having the nerve cell network formation cultivation equipment thing with feature.
When the defiant liquid was introduced to (a) minute description fluidal undercurrent department, to that defiant liquid, the electrode courage where at least part of the courage wall is composed of the weapon porosity material.
Within (b) minute description electrode courage the electrode which formed the chloride NmCl layer of that precious metal to the outer layer department of precious metal Nm is accommodated.
Within (c) minute description electrode courage the electrode solution where chloride NmCl and the alkali metal chloride of the minute description precious metal which charges are solved at gunfire consistency.
[claim5]
5. The 1st claim to there is in which one claim in the 4th claim,
The minute description nerve cell network formation cultivation equipment is used in below (a)-(c) middle which one goal the nerve cell network formation cultivation equipment the fact that with feature.
There is in (a) nerve cell network, measuring the nerve cell ion Channel electric currentUsed to interpretation.
The poem which is a marker of (b) at least Ca imaging interpretation and the former synapse part [nayp] the toe pie shoes or poem [nayp] imaging interpretation, and Endo by MAP2 tagged is a marker of imaging interpretation and winning by providence tagged projection a little or [eyk] the cow a little as for under tagged the low of including used the imaging interpretation by FM1-43 or FM4-64 to imaging interpretation.
Used to high throughput screening system of (c) nerve cell network.
[claim6]
6. There is in a 5th claim,
To the imaging interpretation where the minute description nerve cell network formation cultivation equipment is recorded to (b) as for under using element more than (d)-(f) 1 in the case which is a thing do more the low of having the nerve cell network formation cultivation equipment thing with feature.
Installs the broad dawn equipment which the nerve cell emanates to top of (d) minute description borads.
Installs the investigation equipment which to top of (e) minute description borads investigates the storehouse to the nerve cell or board surface.
The investigation equipment of (f) minute description (e) investigates the storehouse has the condenser for to only the single nerve cell of prescribed.
[claim7]
7. The 1st claim to in the 6th claim the nerve cell network formation cultivation equipment which is recorded to which one claim about under using to under research goal of option the nerve cell network under cultivating at the time of formation as for height as method,
To on minute description even board under (1) cell badge charging nerve cell as for under sowing with process,
By the cell external matrix formation material of (2) cell fixation department and/or from the minute penetration hole of the bottom of cell fixation department sucks the liquid culture medium with every one cell fixation department 1- nerve cell arrangementAt the time of fixation as for height with process,
By the lug of multiple restricts the movement of the nerve cell which became fixed to (3) cell fixation department and by the interval part of the lug of multiple the existence of the nerve cell of the rival each other, synapse junction of the nervous intercellular a chromatic or winning projection recognizes by at the time of formation as for height process as for under including the nerve cell network formation method thing with feature.
[claim8]
8. There is in a 7th claim,
There is in a minute description nerve cell network formation method, when sows the nerve cell from (1) processes, together the glial cell to part other than cell fixation department the low of sowing the nerve cell network formation method thing with feature.
[claim9]
9. There is in a planer patch clamp equipment which used the nerve cell network formation cultivation equipment or this cultivation equipment and the cell badge in on the charging possible even equipment board be about under establishing there is surrounded sows in a equipment borad, with the lug of multiple, the nerve cell to the multiple cell fixation department as the nerve cell sowing device for,
The establishment in on of the minute description equipment board the possible board coat device main body is in the establishment situation, with has together the even bottom to multiple cell fixation department, the lug of multiple is becoming a grade come in contact really, the width which covers a minute description multiple cell fixation department and,
To the device main body (1) nerve cells from the outside supplies the slurry Keane nerve cell suspension at schedule density the suspension supply there to be to an inside of nine for and (2) device main bodies, from minute description suspension supply Guro there is in a terminal of the multiple minute suspension euros which formed continuously with on the basin and (3) minute description each suspension euros, opened to the bottom of the device main body the nerve cell suspension which to minute description each cell fixation department implants formed the suspension injection port for the nerve cell sowing device the fact that with feature.
[claim10]
10. There is in a 9th claim,
There is in a minute description nerve cell sowing device and the multiple minute description suspension euro is set substantially at the identical length, and also, when established the minute description nerve cell sowing device to on the equipment board of the nerve cell network formation cultivation equipment or the planer patch clamp equipment, the minute description suspension injection port of each one accurately to minute description cell fixation department of each one, confronts from the location river course rock is planned the nerve cell sowing device the fact that with feature.
[claim11]
11. The 9th claim or there is in a 10th claim,
The minute description board coat device main body with the upper board which formed a minute description suspension supply nine unites the lower part board which formed a minute description suspension injection port in close adhesion situation, becomes accomplished, the home also, to at least one junction side in the upper board and the lower part board the minute description suspension euro as for under composing forms the nerve cell sowing device the fact that with feature.
[claim12]
12. The 9th claim to there is in which one claim in the 11th claim,
To be a nerve cell network formation cultivation equipment where the minute description nerve cell network formation cultivation equipment is recorded to the 1st claim, and/or is a planer patch clamp equipment where the minute description planer patch clamp equipment is recorded to the 3rd claim the nerve cell sowing device the fact that with feature.
[claim13]
13. The 9th claim to there is in which one claim in the 12th claim,
The minute description device main body is in a equipment board of the minute description nerve cell network formation cultivation equipment or the minute description planer patch clamp equipment, about territory other than minute description cell fixation department implants the minute description nerve cell suspension more has the second suspension euro for the nerve cell sowing device the fact that with feature.
  • Applicant
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • Inventor
  • URISU TSUNEO
  • WANG ZHI HONG
  • UNO HIDETAKA
  • SAITOH MIHO
  • NAGAOKA YASUTAKA
IPC(International Patent Classification)
Reference ( R and D project ) CREST Creation of Nanosystems with Novel Functions through Process Integration AREA
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