WO2020246571A1 - Procédé de production de récipient de culture cellulaire - Google Patents

Procédé de production de récipient de culture cellulaire Download PDF

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Publication number
WO2020246571A1
WO2020246571A1 PCT/JP2020/022234 JP2020022234W WO2020246571A1 WO 2020246571 A1 WO2020246571 A1 WO 2020246571A1 JP 2020022234 W JP2020022234 W JP 2020022234W WO 2020246571 A1 WO2020246571 A1 WO 2020246571A1
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WO
WIPO (PCT)
Prior art keywords
cell
cell culture
recess
diameter
liquid
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/JP2020/022234
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English (en)
Japanese (ja)
Inventor
朋未 牧野
史明 島
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nippon Shokubai Co Ltd
Original Assignee
Nippon Shokubai Co Ltd
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 Nippon Shokubai Co Ltd filed Critical Nippon Shokubai Co Ltd
Priority to JP2021524916A priority Critical patent/JP7368470B2/ja
Publication of WO2020246571A1 publication Critical patent/WO2020246571A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • 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
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus

Definitions

  • An object of the present invention is to provide a method for producing a novel cell culture container, and a method for producing cells and spheroids using the cell culture container obtained by the method.
  • the cell culture substrate has a plurality of recesses having an opening having a diameter of 1000 ⁇ m or less, the inner surface of the recess has a non-cell adhesive surface, and the bottom surface of the recess has cell adhesion.
  • the method according to any one of [1] to [6] above which is a cell culture sheet having a surface.
  • the distance (gap) between the opening and the adjacent opening is not particularly limited, but is, for example, 800 ⁇ m or less, 700 ⁇ m or less, 600 ⁇ m or less, 500 ⁇ m depending on the desired large-scale culture.
  • the range of 300 ⁇ m or less, 200 ⁇ m or less, 100 ⁇ m or less, etc. is exemplified, and any finite value may be used.
  • the material of the cell culture substrate in the present invention is not particularly limited as long as it is a substrate having the above-mentioned recesses on the surface of the substrate. According to the study of the present invention, the material of the base material for cell culture seems to have stronger adhesion of bubbles as the hydrophobicity is higher, but in the present invention, even such a material having high hydrophobicity is convenient. Bubbles can be removed.
  • the material having high hydrophobicity include a fluororesin, a styrene resin (polystyrene, etc.), a polyolefin resin, a polyimide resin, and the like, but the material is not particularly limited and may be a known material.
  • a resin having a fluorine atom in the molecule is preferable, and a fluorine-containing polyimide (fluorine-containing polyimide resin) is more preferable.
  • the polyimide resin used in the present invention is typically obtained by imidizing a polyamic acid obtained by polymerizing one or more kinds of acid dianhydride and diamine.
  • the polyimide resin may contain polyamic acid as part of its chemical structure.
  • As a method for producing the polyimide resin it may be produced by a known method. As an example, the two-stage synthesis method can be used.
  • the divalent organic group represented by X 0 include an alkylene group, an arylene group, an aryleneoxy group, an arylentio group and the like. Further, it may be a condensed ring type divalent hydrocarbon group, a heterocyclic condensed ring type divalent hydrocarbon group, and these oxy groups and thio groups. Among these, an alkylene group, an aryleneoxy group and an arylenthio group are preferable, an alkylene group and an aryleneoxy group are more preferable, and these may be substituted with a fluorine atom.
  • the alkylene group has, for example, 1 to 12 carbon atoms, preferably 1 to 6 carbon atoms.
  • a suitable substituent substituted with a divalent organic group having an aromatic ring is preferably a fluorine atom and / or a trifluoromethyl group, particularly when X 0 does not contain a fluorine atom, and is more preferable. Is a fluorine atom.
  • the polyamic acid When imidizing by thermal imidization, for example, the polyamic acid is placed in air, more preferably in an atmosphere of an inert gas such as nitrogen, helium, or argon, or in vacuum, preferably at a temperature of 50 to 400 ° C. , More preferably 100 to 380 ° C., preferably 0.1 to 10 hours, more preferably 0.2 to 5 hours, to carry out an imidization reaction to obtain a resin composition containing polyimide.
  • an inert gas such as nitrogen, helium, or argon
  • a substance that is more hydrophobic (particularly superhydrophobic) to the resin (and its contact angle)] is particularly preferred, but hydrophilic (particularly superhydrophilic) [eg, hydrophilic or hydrophobic (eg, hydrophobic). ), Or a substance that is more hydrophilic (particularly superhydrophilic) to the cell-adhesive surface or the resin (and the contact angle thereof) constituting the surface is also preferable.
  • hydrophilic particularly superhydrophilic
  • MPC 2-methacryloyloxyethyl phosphorylcholine
  • HEMA hydroxyethyl methacrylate
  • any material known in the art can be used.
  • acrylic resin, silicon resin, synthetic rubber, natural rubber and the like can be mentioned, and a low-elution adhesive layer can be preferably used.
  • Commercially available double-sided tape or the like may be used.
  • the cell culture substrate such as the cell culture sheet thus obtained may be appropriately sized according to the size of the target device from the viewpoint of being used as it is in a known cell culture device.
  • a medium containing cells may be placed on one of the surfaces to carry out cell culture, and the cell culture substrate may be used as a culture plate, each well of the plate, a culture nest (culture dish), a flask, a culture bag, or the like.
  • Cell culture can be carried out on a part or the entire surface of the cell culture substrate which has been housed and fixed in a housing and fixed.
  • room temperature means 20 to 30 ° C.
  • the fluorine-containing polyamic acid resin composition obtained above was applied onto a glass substrate using a die coater so that the thickness of the fluorine-containing polyimide film after firing was 40 ⁇ m to form a coating film. Then, the coating film was fired at 360 ° C. for 1 hour in a nitrogen atmosphere. Then, the fired product was peeled off from the glass substrate to obtain a fluorine-containing polyimide film.
  • the static water contact angle of this fluorine-containing polyimide film was 83.2 °, and the fall angle was 26.4 °.
  • the surface on the PET film side was coated with a spin coater (Mikasa: MS-A150) so that the thickness was 0.05 ⁇ m, and the MPC polymer solution (0.5% ethanol solution) was coated in a dryer at 50 ° C.
  • the time-drying treatment gave a layer having a cell non-adhesive surface [static water contact angle 107.5 ° on the PET film coating layer (MPC polymer coating layer) side].

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Virology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Cell Biology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne un nouveau procédé de production de récipient de culture cellulaire, et un procédé de production de cellules/sphéroïdes utilisant un récipient de culture cellulaire obtenu par ledit procédé de production. La présente invention concerne un procédé de production d'un récipient de culture cellulaire étant pourvu d'un substrat de culture cellulaire comprenant un évidement ayant une ouverture d'un diamètre d'ouverture d'au plus 1000 µm et comprenant un fluide de culture logé au moins dans ledit évidement, le procédé comprenant une étape consistant à distribuer un liquide de distribution à partir d'un orifice de distribution de liquide ayant un diamètre d'ouverture d'au plus 3 mm et/ou possédant un débit d'au moins 100 cm/s; il est préférable que le substrat de culture cellulaire soit une feuille de culture cellulaire étant pourvue d'une pluralité d'évidements ayant chacun une ouverture d'un diamètre d'ouverture d'au plus 1000 µm, et les renfoncements ayant chacun une surface latérale interne comprenant une surface non adhésive de cellule et ayant également chacun un fond comprenant une surface adhésive de cellule.
PCT/JP2020/022234 2019-06-07 2020-06-05 Procédé de production de récipient de culture cellulaire Ceased WO2020246571A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2021524916A JP7368470B2 (ja) 2019-06-07 2020-06-05 細胞培養用容器の製造方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2019107477 2019-06-07
JP2019-107477 2019-06-07

Publications (1)

Publication Number Publication Date
WO2020246571A1 true WO2020246571A1 (fr) 2020-12-10

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ID=73653304

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2020/022234 Ceased WO2020246571A1 (fr) 2019-06-07 2020-06-05 Procédé de production de récipient de culture cellulaire

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JP (1) JP7368470B2 (fr)
WO (1) WO2020246571A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015163043A1 (fr) * 2014-04-22 2015-10-29 株式会社日本触媒 Substrat de culture cellulaire comportant un polymère à base de fluor sur sa surface
WO2015178381A1 (fr) * 2014-05-21 2015-11-26 コニカミノルタ株式会社 Procédé de prétraitement pour dispositif d'étalement de cellules, dispositif d'étalement de cellules, et système de prétraitement pour dispositif d'étalement de cellules
JP2018174824A (ja) * 2017-04-14 2018-11-15 株式会社クラレ マイクロパターンの表面を濡らす方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015163043A1 (fr) * 2014-04-22 2015-10-29 株式会社日本触媒 Substrat de culture cellulaire comportant un polymère à base de fluor sur sa surface
WO2015178381A1 (fr) * 2014-05-21 2015-11-26 コニカミノルタ株式会社 Procédé de prétraitement pour dispositif d'étalement de cellules, dispositif d'étalement de cellules, et système de prétraitement pour dispositif d'étalement de cellules
JP2018174824A (ja) * 2017-04-14 2018-11-15 株式会社クラレ マイクロパターンの表面を濡らす方法

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Publication number Publication date
JP7368470B2 (ja) 2023-10-24
JPWO2020246571A1 (fr) 2020-12-10

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