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WO2019054468A1 - Procédé de libération de cellules d'un échantillon contenant des cellules - Google Patents

Procédé de libération de cellules d'un échantillon contenant des cellules Download PDF

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Publication number
WO2019054468A1
WO2019054468A1 PCT/JP2018/034107 JP2018034107W WO2019054468A1 WO 2019054468 A1 WO2019054468 A1 WO 2019054468A1 JP 2018034107 W JP2018034107 W JP 2018034107W WO 2019054468 A1 WO2019054468 A1 WO 2019054468A1
Authority
WO
WIPO (PCT)
Prior art keywords
bag
container
cell
cells
enzyme
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/JP2018/034107
Other languages
English (en)
Japanese (ja)
Inventor
林　真司
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.)
Kaneka Corp
Original Assignee
Kaneka Corp
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 Kaneka Corp filed Critical Kaneka Corp
Priority to JP2019542298A priority Critical patent/JP7282679B2/ja
Publication of WO2019054468A1 publication Critical patent/WO2019054468A1/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
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/26—Inoculator or sampler

Definitions

  • FIG. 1 An example of the bag for enzyme processing (bag-like container) for performing an enzyme processing of a cell containing sample is shown. It is a cross-sectional schematic diagram for demonstrating the state of the reaction liquid mixture containing a cell containing sample and an enzyme, and the gaseous phase in the bag for enzyme processing.
  • the enzyme processing bag shown in FIG. 1 to which the reaction mixture containing the cell-containing sample and the enzyme and the gas phase (containing material) have been added is sealed with a clamp, and the container of the enzyme processing bag is contained. It is a schematic diagram for demonstrating that the volume of the part to accommodate was adjusted so that it might correspond with the volume of a container.
  • the other example of the bag (bag-like container) for enzyme processing for performing an enzyme processing of a cell containing sample is shown.
  • the other example of the bag (bag-like container) for enzyme processing for performing an enzyme processing of a cell containing sample is shown.
  • the bag-like container used in the present invention is a bag-like container formed of a material having flexibility (also referred to as flexibility). It is preferable that the bag-like container is at least partially made of a transparent or translucent material, because the appearance of the cell suspension inside can be visually observed.
  • the flexible material forming the bag-like container include soft vinyl chloride, vinyl chloride, polyurethane, ethylene-vinyl acetate copolymer, polyolefin such as polyethylene and polypropylene, styrene-butadiene-styrene copolymer Or a thermoplastic elastomer such as a hydrogenated substance thereof and a styrene-isoprene-styrene copolymer or a hydrogenated substance thereof, and a mixture of a thermoplastic elastomer and a softener such as a polyolefin and ethylene-ethyl acrylate, etc.
  • the resin which it has is mentioned.
  • the end portion of the first resin sheet 11 is disposed between the first resin sheet 11 and the second resin sheet 12 on the side of one end in the longitudinal direction of the container main body 10, and the first resin sheet 11 and the second resin sheet 12 around it. Are joined by heat fusion and integrated with the container body 10. Similarly, the tube portion 311 of the first outlet port 31 and the tube portion 321 of the second outlet port 32 have longitudinal ends on the side where the needleless ports 312 and 322 are not disposed.
  • the plan view of the storage portion of the bag-like container is a plan view in a state where the empty bag-like container is spread on a plane so as to widen the area. It is preferable that A / B is the said range in the bag-like container which especially arrange
  • the plan view shape of the storage portion of the bag-like container is not limited to a substantially quadrangle as shown in FIG. 1, and may be any shape such as a circle, an ellipse, a triangle, or a pentagon or more polygon. 1 to 6 show the width A in the width direction and the width B in the longitudinal direction when the entire storage portion 14 is used as the storage portion in the bag-like container 1 whose plan view shape of the storage portion 14 is substantially square.
  • the adipose tissue from 5 persons was treated with collagenase and the steps of preparing the cell suspension under different conditions.
  • a second bag for enzyme treatment (bag-like container 1)
  • two roughly rectangular flexible sheets 11 and 12 of soft polyvinyl chloride having a thickness of 0.4 mm are disposed opposite to each other, and one end side in the longitudinal direction
  • a first inlet port 21, a second inlet port 22, and a third inlet port 23 are provided, and a first outlet port 31 and a second outlet port 32 are provided on the other end side, and the peripheral portion is fused by heat fusion.
  • Forming a fusion-bonded portion 13 to form a bag which is sandwiched between the two sheets 11 and 12 and has a substantially rectangular storage portion 14 having an inner dimension of 67 mm in width and 116 mm in length in an empty state Using.
  • the internal volume of the storage portion 14 of the second enzyme treatment bag is about 120 mL.
  • the fifth bag for enzyme treatment (bag-like container 1) shown in FIG. 6, two approximately rectangular flexible sheets 11 and 12 of soft polyvinyl chloride having a thickness of 0.4 mm are oppositely disposed, and one end in the longitudinal direction
  • the inlet port / outlet port 61 is provided on the side of the sheet, and the peripheral part is fusion-bonded by heat fusion to form the fusion-bonded part 13 so that the two sheets 11 and 12 are empty.
  • the bag is a bag in which a substantially rectangular housing portion 14 having an inner dimension of 18 mm in width and 187.7 mm in length is formed.
  • the internal volume of the container 14 for the fifth enzyme treatment bag is about 22 mL.
  • the outlet port 51 of the third enzyme treatment bag 1 is closed with a lid 313, and 20 mL of adipose tissue and 20 mL of collagenase solution are accommodated in the accommodation portion 14 to form a reaction mixture solution 201, and is accommodated. 3 mL of air is added so that the volume of the air phase 202 in the section 14 becomes 7% of the volume of the accommodation section 14, and the inlet port 41 of the third enzyme treatment bag 1 is sealed with a lid 413 for sealing did.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Selon la présente invention, afin de libérer des cellules nucléées d'un échantillon contenant des cellules tel que des tissus gras avec un traitement enzymatique, le procédé décrit de libération de cellules d'un échantillon contenant des cellules par soumission de l'échantillon contenant des cellules à un traitement enzymatique est caractérisé en ce qu'il comprend une étape d'agitation d'un récipient en forme de sac (1) dans lequel une phase gazeuse (202) et un mélange réactionnel (201) contenant l'échantillon contenant des cellules et une enzyme sont logés et scellés, le rapport du volume de la phase gazeuse (202) étant d'au moins 7 % en volume du volume total de la phase gazeuse (202) et du mélange réactionnel (201).
PCT/JP2018/034107 2017-09-15 2018-09-14 Procédé de libération de cellules d'un échantillon contenant des cellules Ceased WO2019054468A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2019542298A JP7282679B2 (ja) 2017-09-15 2018-09-14 細胞含有試料からの細胞の遊離方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2017-178267 2017-09-15
JP2017178267 2017-09-15

Publications (1)

Publication Number Publication Date
WO2019054468A1 true WO2019054468A1 (fr) 2019-03-21

Family

ID=65722830

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2018/034107 Ceased WO2019054468A1 (fr) 2017-09-15 2018-09-14 Procédé de libération de cellules d'un échantillon contenant des cellules

Country Status (2)

Country Link
JP (1) JP7282679B2 (fr)
WO (1) WO2019054468A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4281131A4 (fr) * 2021-01-19 2025-01-01 Gallant Pet, Inc. Procédés d'optimisation d'une récolte de cellules dérivées de tissu reproducteur et viabilité pour des applications cliniques

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005012480A2 (fr) * 2003-06-25 2005-02-10 Macropore Biosurgery Inc. Systemes et techniques de separation et de concentration de cellules regeneratives prelevees sur des tissus
JP2007185157A (ja) * 2006-01-16 2007-07-26 Kaneka Corp 脂肪組織の酵素処理液から有核細胞を採取する方法
JP2010136628A (ja) * 2008-12-09 2010-06-24 Ihi Corp 細胞培養バッグ及びその細胞培養方法
JP2011078317A (ja) * 2009-10-02 2011-04-21 Olympus Corp 脂肪組織消化装置および細胞分離装置
JP2011172545A (ja) * 2010-02-25 2011-09-08 Olympus Corp 細胞分離装置
JP2015092852A (ja) * 2013-11-12 2015-05-18 倉敷紡績株式会社 生体組織分散用組成物

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6068775A (en) * 1998-04-13 2000-05-30 Circe Biomedical, Inc. Removal of agent from cell suspension
WO2005034843A2 (fr) 2003-09-17 2005-04-21 Macropore Biosurgery Inc. Methodes d'utilisation de cellules regeneratrices dans le traitement d'une maladie vasculaire peripherique et dans d'autres troubles associes
JP2010239891A (ja) 2009-04-03 2010-10-28 Olympus Corp 脂肪組織分離装置
KR20140147805A (ko) 2011-12-07 2014-12-30 아바스템 바이오메디컬 코퍼레이션 지방조직으로부터 비지방 세포의 단리를 위한 방법 및 장치

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005012480A2 (fr) * 2003-06-25 2005-02-10 Macropore Biosurgery Inc. Systemes et techniques de separation et de concentration de cellules regeneratives prelevees sur des tissus
JP2007185157A (ja) * 2006-01-16 2007-07-26 Kaneka Corp 脂肪組織の酵素処理液から有核細胞を採取する方法
JP2010136628A (ja) * 2008-12-09 2010-06-24 Ihi Corp 細胞培養バッグ及びその細胞培養方法
JP2011078317A (ja) * 2009-10-02 2011-04-21 Olympus Corp 脂肪組織消化装置および細胞分離装置
JP2011172545A (ja) * 2010-02-25 2011-09-08 Olympus Corp 細胞分離装置
JP2015092852A (ja) * 2013-11-12 2015-05-18 倉敷紡績株式会社 生体組織分散用組成物

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4281131A4 (fr) * 2021-01-19 2025-01-01 Gallant Pet, Inc. Procédés d'optimisation d'une récolte de cellules dérivées de tissu reproducteur et viabilité pour des applications cliniques

Also Published As

Publication number Publication date
JPWO2019054468A1 (ja) 2020-10-15
JP7282679B2 (ja) 2023-05-29

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