WO2007123035A1 - Incubateur - Google Patents

Incubateur Download PDF

Info

Publication number
WO2007123035A1
WO2007123035A1 PCT/JP2007/057979 JP2007057979W WO2007123035A1 WO 2007123035 A1 WO2007123035 A1 WO 2007123035A1 JP 2007057979 W JP2007057979 W JP 2007057979W WO 2007123035 A1 WO2007123035 A1 WO 2007123035A1
Authority
WO
WIPO (PCT)
Prior art keywords
incubator
side wall
bottom membrane
cell piece
porous
Prior art date
Application number
PCT/JP2007/057979
Other languages
English (en)
Japanese (ja)
Inventor
Keiji Naruse
Norio Ishida
Original Assignee
Strex Incorporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Strex Incorporation filed Critical Strex Incorporation
Priority to JP2008512077A priority Critical patent/JPWO2007123035A1/ja
Publication of WO2007123035A1 publication Critical patent/WO2007123035A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/04—Flat or tray type, drawers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli

Definitions

  • the present invention relates to an incubator for culturing a sample cell piece while generating stress.
  • an incubator formed of a deformable material has a rectangular box shape, and includes a bottom membrane and side walls erected from the entire periphery of the bottom membrane, and a pair of opposing side walls. Is formed with an engaging portion on an extension line of the peripheral edge of the bottom membrane.
  • the bottom membrane is provided with a locking portion, and the cell piece is locked to the locking portion to prevent slippage between the bottom membrane and the cell piece when the incubator is extended.
  • Patent Document 1 International Publication WO2005 / 087913 A1 Publication
  • a first object of the present invention is to propose an incubator capable of applying extremely uniform stress to cell pieces.
  • the inventors of the present invention have intensively studied to achieve the above object, and even if a locking portion is provided on the bottom membrane, it is not possible to completely ensure the integrity of the cell piece and the incubator. I realized I could't.
  • the bottom membrane i.e., the peripheral edge of the cell debris, It was difficult to completely eliminate the bevel.
  • the present inventors have focused on the side wall of the incubator and realized that the present invention should be locked to the periphery of the cell piece. That is, the first aspect of the present invention is defined as follows.
  • An incubator characterized in that the inner surface of the side wall is formed to be porous.
  • the periphery of the cell piece placed on the bottom membrane enters the pores on the inner surface of the side wall, and enters there Locked.
  • the peripheral edge of the cell piece is also extended along with the movement of the side wall thereof, and the peripheral edge of the cell piece does not slide against the bottom membrane of the incubator.
  • the inner surface of the side wall is preferably formed of a foam material. It is an inexpensive and easy-to-control porous material.
  • PDMS Polydimethylsiloxane
  • the substrate of the incubator made of silicon elastomer
  • Any material that has no effect can be selected.
  • the entire circumference of the inner surface of the side wall is porous. However, at least only the inner surfaces of the pair of side walls that are separated from each other when the incubator is extended should be porous. In addition, it is only necessary to make only the portion of the inner surface of the side wall that is in contact with the cell debris porous.
  • a rectangular box type incubator is employed. Making the incubator a rectangular box makes it easier to transport and store incubators that are consumables.
  • the incubator is made of a deformable material. This is because the sample cell piece is indirectly stressed by extending the incubator.
  • a material that does not chemically interfere with the sample cell piece such as silicone elastomer, is used as the incubator forming material.
  • the bottom film has a rectangular shape in plan view, and the entire bottom film is formed to have the same thickness so as to extend uniformly.
  • the bottom membrane is preferably formed of a light-transmitting material so that sample cell fragments can be observed with an optical microscope.
  • the side wall is a thick film to give it mechanical rigidity. This can prevent the bottom film from being randomly deformed.
  • the bottom film and the side wall are preferably integrated from the standpoint of reducing the number of parts and thus the manufacturing cost, but it does not limit the separation of the bottom film and the side wall.
  • Engaging portions are provided on side walls provided at a pair of opposing edges of the rectangular bottom film. This engaging portion may be engaged with the fixed plate and the moving plate of the extension device. Thus, the change in position of the two members whose relative positions can be controlled causes the incubator to be deformed.
  • the direction of the stress exerted on the sample cell piece on the deformed portion is different from those in other portions. Therefore, it is preferable to form the engaging portion on the extended line of the peripheral edge in the extending direction of the rectangular bottom film. As a result, the force applied to the engaging portion is applied to the peripheral edge in the extending direction of the bottom membrane, and this is reliably pulled. Therefore, the deformation of the peripheral edge is prevented, and uniform stress can be applied to all the sample cell pieces on the bottom membrane.
  • vascular endothelial cells such as human 'monkey pig' rush 'rat' mouse 'rabbit
  • smooth muscle cells cardiomyocytes, skeletal muscle cells, fibroblasts, osteoblasts, chondrocytes, osteoclasts, A nerve cell etc.
  • FIG. 1 is a perspective view of an incubator according to an embodiment of the present invention.
  • FIG. 2 shows an incubator according to an embodiment of the present invention
  • (A) is a plan view
  • (B) is a front view
  • (C) is a bottom view
  • (D) is a right side view.
  • FIG. 3 is a cross-sectional view taken along line AA in FIG.
  • FIG. 4 is a cross-sectional view showing how the incubator of the example is used.
  • FIG. 5 is a photograph showing a PDMS foaming barter.
  • Fig. 6 is an enlarged photograph of the surface of PDMS foaming barta.
  • FIG. 7 is a surface electron micrograph of PDMS foaming barta.
  • FIG. 1 shows an incubator 21 of this example
  • FIG. 1 is a perspective view thereof
  • FIG. 2 (A) is a plan view
  • FIG. 2 (B) is a front view
  • FIG. 2 (C) is a bottom view
  • FIG. D) is a right side view.
  • FIG. Fig. 3 is a cross-sectional view taken along line AA in Fig. 2 (A).
  • Figure 4 is a usage diagram.
  • the incubator 21 of the embodiment is a box shape molded with a transparent silicone elastomer, and includes a thin bottom film 23 and side walls 25 and 26 erected integrally from the periphery of the bottom film 23.
  • the bottom film 23 has a thickness of about 100 ⁇ m to about 200 ⁇ m
  • the side wall 25 has a thickness of about 1 cm
  • the side wall 26 has a thickness of about 2 mm.
  • An engaging hole 27 is formed in the side wall 25.
  • Fibronectin, collagen, etc. are applied to the surface of the bottom membrane 23 in order to implant cell debris.
  • a protrusion 31 is formed as a locking portion from the bottom film 23.
  • the protrusion 31 interferes with the sample cell piece 29 and prevents slippage between the protrusion 31 and the bottom film 23. Accordingly, uniform stress can be applied to the sample cell piece 29.
  • irregularities can be formed on the surface of the bottom film 23. The unevenness and protrusion 31 can be omitted.
  • Porous bodies 41, 41 are laminated on the inner surfaces of the side walls 25, 25, and many The porous bodies 43 and 43 are laminated.
  • These porous bodies 43 are formed as follows.
  • PDMS containing the above water was poured into a hot plate (ASONE, ND-1) heated to 100 ° C.
  • the PDMS was heated to 100 ° C for 15-30 minutes with a thickness of about 0.5-2 mm.
  • PDMS polymerized while foaming.
  • Fig. 5 shows a picture of the PDMS foaming barta removed from the hot plate.
  • Fig. 6 shows an enlarged photograph of the surface of this foamed barta. From Fig. 6, a depression of several millimeters can be confirmed on the surface. Cell debris is locked to the peripheral wall of this depression.
  • Fig. 7 shows a micrograph of the surface. From Fig. 7, it can be seen that the holes in the foamed butter are in communication, and that each hole has a smaller diameter as it goes into the inside where the diameter is larger on the surface side.
  • the PDMS foaming barter thus obtained is cut into a predetermined shape to form porous bodies 41, 43.
  • the porous bodies 41 and 43 are bonded to the inner surfaces of the side walls 25 and 26 using a general-purpose adhesive.
  • FIG. 4 shows an extension device 50 according to the embodiment.
  • the fixed plate 51 is fixed to the rail plate 56
  • the movable plate 53 is slidably mounted on the rail plate 56.
  • Pins 52 and 54 project from the fixed plate 51 and the movable plate 53, respectively, and are inserted into the engagement holes 27 of the incubator 21.
  • the movable plate 53 moves through the rod in the direction of the arrow as the step motor 55 rotates.
  • Reference numeral 57 denotes a control device that controls the rotation of the step motor 55. In the embodiment, a computer device is used.
  • the engagement hole 27 is located on the extended line of the peripheral edges 24, 24 of the bottom film 23. More preferably, as shown in the drawing, the extension line coincides with the outer edge portion of the engagement hole 27. As a result, the force from pin 32 to pin 34 is more directly directed to the peripheral edge 24 of the bottom membrane 23. It will take. Therefore, the depression of the peripheral edge 24 is prevented, and the bottom film 23 is uniformly extended. Therefore, the stress applied to the sample cell piece 29 on the bottom film 23 becomes uniform.
  • the porous bodies 41 and 43 are attached to the inner surfaces of the peripheral walls 25 and 26, the periphery of the cell piece 29 is locked to the porous bodies 41 and 43.
  • the cell pieces 29 are also extended along the porous bodies 41 and 43 and do not slide against the bottom membrane 23 of the incubator 21.
  • the protrusion 31 is formed on the bottom film 23, the slip of the cell piece 29 with respect to the bottom film 23 is more reliably prevented.
  • the rectangular side wall incubator has a porous side wall, but the shape of the incubator is not particularly limited.
  • a side wall may be provided on a culture membrane that is deformed by suction, and the inner surface thereof may be made porous.

Landscapes

  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Clinical Laboratory Science (AREA)
  • Immunology (AREA)
  • Mechanical Engineering (AREA)
  • Cell Biology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention concerne un incubateur dans lequel une contrainte peut être uniformément appliquée à un élément de cellule. L'incubateur ayant une forme de type boîte rectangulaire, formée à partir d'une matière déformable, comprend une membrane inférieure et des parois latérales installées de façon à s'étendre verticalement à partir de la bordure périphérique entière de la membrane inférieure. Les surfaces internes des parois latérales sont rendues poreuses.
PCT/JP2007/057979 2006-04-18 2007-04-11 Incubateur WO2007123035A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2008512077A JPWO2007123035A1 (ja) 2006-04-18 2007-04-11 培養器

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2006-114941 2006-04-18
JP2006114941 2006-04-18

Publications (1)

Publication Number Publication Date
WO2007123035A1 true WO2007123035A1 (fr) 2007-11-01

Family

ID=38624940

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2007/057979 WO2007123035A1 (fr) 2006-04-18 2007-04-11 Incubateur

Country Status (2)

Country Link
JP (1) JPWO2007123035A1 (fr)
WO (1) WO2007123035A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008271964A (ja) * 2007-03-30 2008-11-13 Strex Inc 受精卵の培養方法及び受精卵の培養装置
WO2012147878A1 (fr) * 2011-04-27 2012-11-01 株式会社メニコン Récipient de culture cellulaire
JP2013255494A (ja) * 2012-06-13 2013-12-26 sheng-nan Chang 生体力学的負荷測定用容器
WO2019069708A1 (fr) * 2017-10-05 2019-04-11 ソニーセミコンダクタソリューションズ株式会社 Dispositif de détection de potentiel électrique de cellule, procédé de fabrication de dispositif de détection de potentiel électrique de cellule et système de traitement d'informations

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH10155475A (ja) * 1996-12-02 1998-06-16 Toru Takemasa シリコンベルトを使った培養細胞用伸縮刺激負荷装置
WO2004085606A1 (fr) * 2003-03-24 2004-10-07 National Institute For Environmental Studies Milieu de culture cellulaire et preparation solidifiee de proteine ou de peptide d'adhesion cellulaire

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH10155475A (ja) * 1996-12-02 1998-06-16 Toru Takemasa シリコンベルトを使った培養細胞用伸縮刺激負荷装置
WO2004085606A1 (fr) * 2003-03-24 2004-10-07 National Institute For Environmental Studies Milieu de culture cellulaire et preparation solidifiee de proteine ou de peptide d'adhesion cellulaire

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008271964A (ja) * 2007-03-30 2008-11-13 Strex Inc 受精卵の培養方法及び受精卵の培養装置
WO2012147878A1 (fr) * 2011-04-27 2012-11-01 株式会社メニコン Récipient de culture cellulaire
JPWO2012147878A1 (ja) * 2011-04-27 2014-07-28 株式会社メニコン 細胞培養器
JP6074860B2 (ja) * 2011-04-27 2017-02-08 国立大学法人 岡山大学 細胞培養器
JP2013255494A (ja) * 2012-06-13 2013-12-26 sheng-nan Chang 生体力学的負荷測定用容器
WO2019069708A1 (fr) * 2017-10-05 2019-04-11 ソニーセミコンダクタソリューションズ株式会社 Dispositif de détection de potentiel électrique de cellule, procédé de fabrication de dispositif de détection de potentiel électrique de cellule et système de traitement d'informations
US11584909B2 (en) 2017-10-05 2023-02-21 Sony Semiconductor Solutions Corporation Cell potential detection device, method of manufacturing cell potential detection device, and information processing system

Also Published As

Publication number Publication date
JPWO2007123035A1 (ja) 2009-09-03

Similar Documents

Publication Publication Date Title
WO2005087913A1 (fr) Dispositif de culture
Wang et al. Fabrication of well-defined mushroom-shaped structures for biomimetic dry adhesive by conventional photolithography and molding
Yi et al. Simple and reliable fabrication of bioinspired mushroom-shaped micropillars with precisely controlled tip geometries
Xue et al. Reversible adhesion switching of porous fibrillar adhesive pads by humidity
Yi et al. Continuous and scalable fabrication of bioinspired dry adhesives via a roll-to-roll process with modulated ultraviolet-curable resin
US10052811B2 (en) Wrinkled surfaces with tunable hierarchy and methods for the preparation thereof
KR101174771B1 (ko) 소프트 몰드 및 이의 제조방법
WO2007123035A1 (fr) Incubateur
Chidambaram et al. High fidelity 3D thermal nanoimprint with UV curable polydimethyl siloxane stamps
DE60228065D1 (de) Verfahren zur herstellung von mikronadelstrukturen mit sanfter lithographie und photolithographie
RU2695290C2 (ru) Способ изготовления штампа с рисунком, штамп с рисунком и способ отпечатывания
Shah et al. Micropatterning of proteins and mammalian cells on indium tin oxide
US20160160165A1 (en) Methods, apparatuses, and systems for cell and tissue culture
Bhandaru et al. Lithographic tuning of polymeric thin film surfaces by stress relaxation
Lopera et al. Plasma‐Based Surface Modification of Polydimethylsiloxane for PDMS‐PDMS Molding
Shen et al. UV‐embossed microchannel in biocompatible polymeric film: application to control of cell shape and orientation of muscle cells
KR101222946B1 (ko) 백 프레인이 부착된 소프트 몰드의 제조방법
Bhandaru et al. Programmable nanopatterns by controlled debonding of soft elastic films
Chan-Park et al. Simulation and investigation of factors affecting high aspect ratio UV embossing
Lee et al. On-chip parylene-C microstencil for simple-to-use patterning of proteins and cells on polydimethylsiloxane
CN103814428A (zh) 超疏水薄膜片材的制备方法
Murray et al. Fabrication of polymeric substrates with micro-and nanoscale topography bioimprinted at progressive cell morphologies
WO2005090548A1 (fr) Dispositif de maintien pour incubateur
CN101487971A (zh) 软模及其制造方法
KR102159231B1 (ko) 세포 배양용 용기 및 이의 제조방법

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 07741415

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2008512077

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 07741415

Country of ref document: EP

Kind code of ref document: A1