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Formation and use of neuronal network, and neuron seeding device 新技術説明会

外国特許コード F140007985
整理番号 AF19-04TW
掲載日 2014年10月7日
出願国 台湾
出願番号 102112687
公報番号 201413603
公報番号 I598830
出願日 平成25年4月10日(2013.4.10)
公報発行日 平成26年4月1日(2014.4.1)
公報発行日 平成29年9月11日(2017.9.11)
優先権データ
  • 特願2012-205561 (2012.9.19) JP
発明の名称 (英語) Formation and use of neuronal network, and neuron seeding device 新技術説明会
発明の概要(英語) (TWI598830)
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.
特許請求の範囲(英語) [claim1]
1. Kinds of nerve cell networks form with the raise installment, its characteristic is on may the complementary cell culture medium smooth foundation plate, forms by the cell stators of many prominence department surrounding, this cell stator meets the following (1)~(3) claim, (1) in many prominence departments of constitution cell stator, establishes the wide gap, by does not enable the nerve cell the cell body to pass; More than (2) is may hold 1~ several nerve cells because of the cell stator's inside diameter that prominence department institutes stipulated the size of cell body; (3) constitution cell stator the foundation plate surface of bottom surface's has in following (A) and (B) at least essential factor; the (A) superposition has the extracellular matrix to form material, the tiny birthplace through hole that (B) is establishes the culture medium of attraction that attraction installment uses because of the above foundation plate surface, and is equipped with does not cause the aperture who the nerve cell adopts.
[claim2]
2. According to claim 1, nerve cell network forms with the raise installment, the above raise installment conforms to below in (1)~(3) any one item, (1) forms the achievement on the above foundation plate for the choice region's cell stator, disposes 1~ several achievement in this cell stator for the choice cell's nerve cell, simultaneously other nerve cells is only sowing seeds on the foundation plate; (2) forms the plural number cell stator by the suitable mutual gap on the above foundation plate, disposes 1~ several nerve cells in these cell stators, simultaneously 1 cell stator will use as the disposition nerve cell's choice region; (3) by may form the plural number on the above foundation plate either most quite Yu Shangshu (1) or (2) nerve cell network unit way, causes the above cell stator to disperse in the high flux analysis that of nerve cell network the suitable position forms with the raise installment.
[claim3]
3. According to claim 1, or the 2nd nerve cell network forms with the raise installment, the above nerve cell network forms with the raise installment is takes the nerve cell network as the plane formula of piece membrane immobilization installment object, (1) above foundation plate for the electric insulation foundation plate, the establishment enables to constitute this electric insulation foundation plate cell stator the foundation plate surface both sides surface connections of bottom surface the above tiny birthplace through hole, (2) tiny birthplace through hole's first superficial side's nerve cell network forms the side and with its opposite side second superficial side, separately establishment with taking maintaining as above cell culture medium storing of conductive liquidThe fluid department and for may to store fluid department the electricity pole piece of conductive liquid circular telegram with the disposition, (3) first superficial side's storing fluid department served as to fix in the storing fluid department that the nerve cell of above cell stator uses.
[claim4]
4. According to claim 3, nerve cell network forms with the raise installment, in the above plane formula piece membrane immobilization installment, the above first superficial side and second superficial side's electricity pole piece has the following (a)~(c) essential factor, when (a) above stores in the fluid department inducts the conductive liquid, contacts this conductive liquid at least sub-system of vessel wall by the electrode vessel that the inorganic porous material is constituted; the (b) accommodation in the precious metal Nm surface layer department forms this precious metal in the above electrode vessel the chloride NmCl level electrode; (c) fills in the above electrode vessel has above precious metal chloride NmCl and alkaline metal chloride electrode solution by the saturated density dissolution.
[claim5]
5. Like forms the application patent scope 1st item to the 4th any nerve cell network with the raise installment, the above nerve cell network forms with the raise installment is the use any goal in following (A)~(C), (A) use nerve cell ion channel electric current the survey in nerve cell network and analysis of; the (B) use at least contains the Ca image formation analysis, as well as, outside secretes body's FM1-43 or FM4-64 the image formation analysis of image formation analysis because of the mark inner body because of former synapse spot tracer synapse element (Synaptophysin) either the image formation analysis and because of tree suddenly tracer MAP2 mark the image formation analysis of synapse protein (Synapsin) mark; the (C) use screens the system in the nerve cell network's high flux.
[claim6]
6. According to claim 5, nerve cell network forms with the raise installment, when the above nerve cell network forms with the raise installment is the use in the image formation analysis of (B) record, has further in following (D)~(F) 1 above essential factor, (D) above foundation plate's upside, the establishment bright after the light that the cell sends out was installed by the light; (E) above foundation plate's upside, the establishment has or foundation plate surface illumination light the irradiation unit to the nerve cell; the (F) above (E) irradiation unit provides is useful in only to assigning the sole nerve cell illumination light condensation system.
[claim7]
7. 1 nerve cell network methods of formation, it is the use like forms the application patent scope 1st item to the 6th any nerve cell network with the raise installment, forms under the raise based on the random research goal the nerve cell network method, its characteristic is includes: (1) on the above smooth foundation plate under cell culture medium packing, sowing seeds nerve cell's step; And (2) forms the material because of the extracellular matrix of cell stator and/or because of from cell stator the tiny birthplace through hole attraction liquid media of bottom surface, in each cell stator, disposition/fixed 1~ several nerve cells' steps; And (3) fixes because of the plural number prominence department limit in cell stator the migration of nerve cell, simultaneously breaks out the gap part because of plural number, makes to know mutually the opposite party nerve cell existence, forms use axon or tree nerve intercellular space's the step of synapse joint suddenly.
[claim8]
8. According to claim 7, nerve cell network method of formation, when above nerve cell network method of formation, in (1) step sowing seeds nerve cell, one and neuroglia cell sowing seeds part outside cell stator.
[claim9]
9. 1 kinds of nerve cell seed attachments, it is the establishment forms in the nerve cell network with the raise installs or uses in the plane formula piece membrane immobilization installment of this raise installment may the complementary cell culture medium smooth installment foundation plate on, for was installed nerve cell sowing seeds of nerve cell seed attachment that many cell stators the plural number prominence department institute in foundation plate surrounds, its characteristic is may establish on above installment foundation plate the plate-type installment main body, has the cover above most cell stators' sizes under its establishment condition, simultaneously its smooth bottom surface's in many with most cell stators breaks out the peak contact, the installment main body establishes to have(1) is used from the exterior supplies by certain density aerosol nerve cell's nerve cell suspending liquid suspending liquid supplies mouth, (2) in the installment main body interior, extends most tiny aerosol liquid flow roads of establishment by the above suspending liquid supplies mouth ingredient branched, and (3) in various above aerosol liquid flow roads' terminals, opens the mouth it to pour into the nerve cell suspending liquid to the installment main body's bottom surface various above cell stators' suspending liquid filling openings.
[claim10]
10. According to claim 9, nerve cell seed attachment, in the above nerve cell seed attachment, most above aerosol liquid flow roads materially establish the same length, and when the nerve cell network forms with raising the equipment or on the installment foundation plate of plane formula piece membrane immobilization installment establishes the above nerve cell vaccination installment, each above suspending liquid filling opening is designed correctly to correspond to each above cell stator locates.
[claim11]
11. According to claim 9, or 10 nerve cell seed attachments, above plate-type installment this system will establish to have the above board and establishment of above suspending liquid supplies mouth has the below board of above suspending liquid filling opening by being closed condition joint, and in the upside board with a below board's at least adjoining plane, forms the constitution above aerosol liquid flow road's trench.
[claim12]
12. According to claim 9, item to the 11th any nerve cell seed attachment, the above nerve cell network forms with the raise installment is according to claim 1, nerve cell network forms with the raise installment, and/or the above plane formula piece membrane immobilization installment is according to claim 3, plane formula piece membrane immobilization installment.
[claim13]
13. According to claim 9, item to the 12th any nerve cell seed attachment, the above installment main body has further for forms to the above nerve cell network with the raise installment or above plane formula piece membrane immobilization installment installment foundation plate the region in outside above cell stator, injects 2nd the aerosol liquid flow road system above nerve cell suspending liquid.
  • 出願人(英語)
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • 発明者(英語)
  • URISU TSUNEO
  • WANG ZHI-HONG
  • UNO HIDETAKA
  • NAGAOKA YASUTAKA
  • SAITO MIHO
国際特許分類(IPC)
参考情報 (研究プロジェクト等) CREST Creation of Nanosystems with Novel Functions through Process Integration AREA
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