WO2014131846A1 - Insert de culture, dispositif de culture pour cellules, cultures de tissus ainsi que levure et bactéries - Google Patents

Insert de culture, dispositif de culture pour cellules, cultures de tissus ainsi que levure et bactéries Download PDF

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Publication number
WO2014131846A1
WO2014131846A1 PCT/EP2014/053857 EP2014053857W WO2014131846A1 WO 2014131846 A1 WO2014131846 A1 WO 2014131846A1 EP 2014053857 W EP2014053857 W EP 2014053857W WO 2014131846 A1 WO2014131846 A1 WO 2014131846A1
Authority
WO
WIPO (PCT)
Prior art keywords
holder
membrane
legs
cultivating
cells
Prior art date
Application number
PCT/EP2014/053857
Other languages
German (de)
English (en)
Inventor
Hans Nagels
Simone Valley
Original Assignee
Alpha Plan Gmbh
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 Alpha Plan Gmbh filed Critical Alpha Plan Gmbh
Publication of WO2014131846A1 publication Critical patent/WO2014131846A1/fr

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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
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
    • 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
    • C12M23/14—Bags
    • 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
    • C12M23/42—Integrated assemblies, e.g. cassettes or cartridges
    • 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
    • C12M23/46—Means for fastening
    • 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
    • C12M25/02—Membranes; Filters

Definitions

  • the invention relates to culturing inserts for cells and tissue cultures as well as yeasts and bacteria as objects and devices for cultivating cells and tissue cultures as well as yeasts and bacteria as objects with cultivating inserts.
  • cells are grown by standard on polystyrene surfaces. This is done, for example, in sterile Petri dishes or so-called T-bottles.
  • inserts are known whose bottom consists of a membrane. These have different sized pores, whereupon cells are cultivated. These cell culture inserts are used for so-called co-cultivations.
  • co-cultivations In addition to the cells on the membrane of the inserts, there are cells on the bottom of the vessel in which the cell culture insert hangs or stands.
  • the document DE 101 28 810 A1 discloses a device for cultivating cells, in particular human or animal cells.
  • cells of at least one specific type each have a culture set in a defined environment, the cells of the relevant culture being supplied with associated liquid nutrient media, growth factors, gases or the like.
  • a predetermined number of cell culture chambers are provided, which can be connected either in series or in parallel.
  • the cell culture chambers are self-contained as chambers. A special design is not provided.
  • Document DE 696 17 303 T2 discloses a two-tank cell culture insert in which the containers are separated by a microporous membrane.
  • the cell culture insert is first placed horizontally in the media container and then placed vertically. Only then can the cultivation take place in the first chamber. Before the second cultivation in the second chamber either the medium must be completely changed or the medium in the first chamber to be aspirated and filled with new medium. Only then does the first cultivation take place. It is not excluded that one of the two membrane sides can come into contact with the medium, which is intended for the other side of the membrane.
  • Document US 2004/0043 481 A1 describes a device and a method for cultivating and maintaining cells and tissues in which the risk of contamination is to be minimized. In most cases, however, only one-sided cultivation is possible.
  • a reversible membrane insert for the cultivation of cells is known.
  • On a frame a membrane is stretched. Around the frame is an O-ring for sealing.
  • the frame is inserted into a housing.
  • the exact position of the frame in the housing can be determined.
  • Through recesses in the lower region of the housing medium can penetrate when the housing is placed in an initially empty medium container and then medium is filled on the outside of the housing in the medium container. From the top then a second medium can be applied.
  • the housing including the frame is rotated and a new medium and a new cell culture can be applied from above.
  • air bubbles can not escape completely through the recesses. This is only possible if the housing is dipped obliquely into the medium. As a result, however, the housing can not be used in custom-fit containers.
  • the invention disclosed in claims 1 and 8 has for its object to provide a simple cultivation use for cells, tissue cultures, yeasts and bacteria.
  • the cultivation inserts and the devices for cultivating cells and tissue cultures as well as yeasts and bacteria with such cultivating inserts are characterized in particular by their simple realization with simultaneous ease of handling.
  • first annular and a first chamber-forming holder with first legs and a pore-containing membrane and thus culturable from both sides, so that the membrane spans the interior of the first holder
  • the object to be cultivated is simply placeable on the membrane of the first annular holder.
  • the second annular holder forms a second chamber, wherein the membrane is the bottom of this second chamber.
  • the second annular holder is displaceable relative to the first annular holder.
  • the first annular holder with the membrane with the objects forms a primary side which can be contacted with a medium.
  • the first annular support with the membrane as the bottom can now be provided with a cell culture insert as a secondary side.
  • the first annular support forms the first chamber for this purpose.
  • the membrane has pores, for example, with a size of 3 to 15 ⁇ .
  • the membrane may be designed to disk or cup-shaped.
  • the disc-shaped membrane can have depressions for at least one first object in each case.
  • a depression may have a base area of, for example, less than or equal to 50 ⁇ 50 ⁇ m 2 .
  • the annular holders have legs. This leads to air being able to escape, thus ensuring complete wetting of the membrane and of the cells arranged thereon.
  • the legs of the annular holder advantageously result in that the air can escape between the membrane and the bottom of the body when filling the body.
  • the membrane on the first holder can advantageously terminate with the edge regions of the second holder between the second legs or project beyond these edge regions.
  • the objects on the primary ropes can thus be completely supplied with the medium in the body. Bubbles caused by air bubbles during wetting are avoided.
  • the voided membrane causes objects to permeate the membrane during culture and affect objects on the secondary side of the membrane.
  • the first annular holder projects beyond the first legs, so that a stop for the latching noses is present.
  • end portions of the first legs are designed as stops for the latching noses according to the embodiment of claim 3.
  • the holder can be easily moved against each other, without causing the holders are separated from each other due to the existing in both directions of movement stops.
  • the membrane according to the embodiment of claim 4 consists of a polycarbonate or polyethylene terephthalate.
  • the membrane has according to the embodiment of claim 5 pores equal to / greater than 0.1 ⁇ and less than / equal to 15 ⁇ .
  • the membrane is a nuclear track membrane.
  • the membrane is according to the embodiment of claim 7, glued or sealed to the first holder membrane. Gluing and sealing are known joining techniques that ensure a secure hold of the membrane on the holder.
  • An embodiment of the invention is illustrated in principle in the drawings and will be described in more detail below.
  • FIG. 1 shows a cultivating insert for cells and tissue cultures as well as yeasts and bacteria as objects in an exploded view
  • FIG. 3 shows a device with cultivation inserts.
  • a culture use for cells 7 and tissue cultures and yeasts and bacteria as objects essentially consists of a first holder 1 with first legs 2, a membrane 3, a second holder 4 with second legs 5 and an O-ring seal 6.
  • Fig. 1 shows a cultivation use for cells and tissue cultures and yeasts and bacteria as objects in a schematic exploded view.
  • the first holder 1 is a first annular and a first chamber forming holder 1 with the first legs 2.
  • the membrane 3 is applied.
  • the membrane 3 spans the interior of the first holder 1 and thus forms a bottom.
  • the membrane may consist of a polycarbonate or polyethylene terephthalate.
  • the membrane 3 has pores, so that the membrane is a membrane 3 cultivable from both sides. The pores are, for example, equal to / greater than 0.1 ⁇ and less than or equal to 10 ⁇ .
  • the second holder 4 is an annular and a second chamber forming holder 4 for the first holder. 1 Furthermore, the second holder 4 has second legs 5, so that the first legs 2 and the second legs 5 face outward.
  • the O-ring seal 6 is arranged sealingly, which rests against the outer surface of the first Garis 1 and the inner surface of the second holder 4.
  • the first holder 1 in the second holder 4 is displaceable.
  • the first holder 1 have a circumferential groove for the O-ring seal 6.
  • FIGS. 2a to 2c show a cultivating insert with objects in basic representations.
  • FIG. 3 shows a device with cultivation inserts in a basic representation.
  • the device for cultivating cells and tissue cultures and yeasts and bacteria as objects consists of a trough-shaped and filled with a medium 9 body 10 in which several cultivation inserts are used.
  • the assembled cultivation insert is inserted into the trough-shaped body 10.
  • 4 ml of medium 9 with first objects 7 are applied to the membrane 3.
  • the second holder 4 forms a chamber with the membrane 3.
  • the first cells 7 grow on the primary side thus present (representation in FIG. 2 a).
  • the spent medium 9 is removed after a time determined by cell growth.
  • the second holder 4 is now pressed down (representation in FIG. 2b).
  • the cultivation insert is rotated by 180 ° (shown in Fig. 2c).
  • fibroblasts with collagen are given as second objects 8 on the secondary side and covered with medium 1 1.
  • medium 9 is placed in the trough-shaped body 10, so that the membrane 3 with the adhering first objects 7 are on the primary side in the medium 9 and the primary side has air bubble-free contact with the medium 9. Now the device with the cultivation inserts can be transferred to an incubator for further incubation.
  • 4 locking lugs are located on the inner surface of the second holder. Furthermore, the first annular holder 1 projects beyond the first legs 2, so that a stop for the latching noses is present. In addition, the end portions of the first legs 2 are designed as stops for the latching noses.

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  • 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)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Clinical Laboratory Science (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention concerne des inserts de culture pour cellules, cultures de tissus ainsi que levures et bactéries en tant qu'objets, et des dispositifs dotés d'inserts de culture. Les inserts de culture et les dispositifs se distinguent notamment par leur mode de réalisation simple tout en étant faciles à manipuler. A cet effet, un insert de culture présente un premier support qui est de forme annulaire et qui forme une première chambre dotée de premiers pieds et d'une membrane qui présente des pores et est ainsi cultivable des deux côtés, de telle façon que la membrane recouvre l'espace interne du premier support; un deuxième support qui est de forme annulaire, qui forme une deuxième chambre pour le premier support, et qui est doté de deuxièmes pieds, de telle façon que les premiers pieds et les deuxièmes pieds soient orientés vers l'extérieur; et un joint torique disposé sur le face externe du premier support et sur la face interne du deuxième support, de telle façon que le joint torique se trouve entre le premier support et le deuxième support et que le premier support puisse puise se déplacer dans le deuxième support.
PCT/EP2014/053857 2013-02-27 2014-02-27 Insert de culture, dispositif de culture pour cellules, cultures de tissus ainsi que levure et bactéries WO2014131846A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102013203306.0A DE102013203306B4 (de) 2013-02-27 2013-02-27 Einsatz zur beidseitigen Kultivierung einer Poren aufweisenden Membran für Zellen und Gewebekulturen sowie Hefen und Bakterien als Objekte
DE102013203306.0 2013-02-27

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WO2014131846A1 true WO2014131846A1 (fr) 2014-09-04

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Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108144559A (zh) * 2018-02-24 2018-06-12 清华大学 一种基于核径迹膜的催化膜反应器及其制备方法
US11608486B2 (en) 2015-07-02 2023-03-21 Terumo Bct, Inc. Cell growth with mechanical stimuli
US11613727B2 (en) 2010-10-08 2023-03-28 Terumo Bct, Inc. Configurable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11624046B2 (en) 2017-03-31 2023-04-11 Terumo Bct, Inc. Cell expansion
US11629332B2 (en) 2017-03-31 2023-04-18 Terumo Bct, Inc. Cell expansion
US11634677B2 (en) 2016-06-07 2023-04-25 Terumo Bct, Inc. Coating a bioreactor in a cell expansion system
US11667881B2 (en) 2014-09-26 2023-06-06 Terumo Bct, Inc. Scheduled feed
US11667876B2 (en) 2013-11-16 2023-06-06 Terumo Bct, Inc. Expanding cells in a bioreactor
US11685883B2 (en) 2016-06-07 2023-06-27 Terumo Bct, Inc. Methods and systems for coating a cell growth surface
US11795432B2 (en) 2014-03-25 2023-10-24 Terumo Bct, Inc. Passive replacement of media
US11965175B2 (en) 2016-05-25 2024-04-23 Terumo Bct, Inc. Cell expansion
US11999929B2 (en) 2020-04-10 2024-06-04 Terumo Bct, Inc. Methods and systems for coating a cell growth surface

Families Citing this family (1)

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Publication number Priority date Publication date Assignee Title
DE102017213923A1 (de) * 2017-08-10 2019-02-14 Robert Bosch Gesellschaft Für Medizinische Forschung Mbh Vorrichtung zur Kultivierung von Gewebeschnitten

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EP0700990A2 (fr) * 1994-09-12 1996-03-13 Becton, Dickinson and Company Bouteilles tournantes pour la co-culture trans-membrane de cellules et son procédé d'utilisation
DE69126882T2 (de) * 1990-10-10 1997-11-20 Costar Corp Kulturvorrichtung mit lösbarer aufwuchsfläche für zellen oder gewebe
DE69617303T2 (de) * 1995-03-30 2002-07-11 Becton Dickinson Co Transmembran Ko-Kultureinsatz und Verfahren zu deren Anwendung
DE69624483T2 (de) * 1995-08-03 2003-06-18 Corning Inc Umkehrbarer membraneinsatz zur kultivierung von zellen
WO2006131123A2 (fr) * 2005-06-10 2006-12-14 Nunc A/S Support d'element d'insertion de culture, element d'insertion de culture et systeme d'elements d'insertion de culture
WO2010042072A1 (fr) * 2008-10-08 2010-04-15 Agency For Science, Technology And Research Appareil pour la culture de cellules ancrage-dépendantes

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US5665596A (en) * 1995-07-31 1997-09-09 Becton, Dickinson And Company Device for cell co-culture and method for its use in culturing cells
DE10128810B4 (de) 2001-06-18 2013-11-07 Pan-Biotech Gmbh Einrichtung zur Kultivierung von Zellen, insbesondere menschlicher oder tierischer Zellen
US7229820B2 (en) * 2002-06-13 2007-06-12 Wilson Wolf Manufacturing Corporation Apparatus and method for culturing and preserving tissue constructs

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE69126882T2 (de) * 1990-10-10 1997-11-20 Costar Corp Kulturvorrichtung mit lösbarer aufwuchsfläche für zellen oder gewebe
US5470743A (en) * 1991-03-06 1995-11-28 Becton, Dickinson And Company Transmembrane cell culture device
EP0700990A2 (fr) * 1994-09-12 1996-03-13 Becton, Dickinson and Company Bouteilles tournantes pour la co-culture trans-membrane de cellules et son procédé d'utilisation
DE69617303T2 (de) * 1995-03-30 2002-07-11 Becton Dickinson Co Transmembran Ko-Kultureinsatz und Verfahren zu deren Anwendung
DE69624483T2 (de) * 1995-08-03 2003-06-18 Corning Inc Umkehrbarer membraneinsatz zur kultivierung von zellen
WO2006131123A2 (fr) * 2005-06-10 2006-12-14 Nunc A/S Support d'element d'insertion de culture, element d'insertion de culture et systeme d'elements d'insertion de culture
WO2010042072A1 (fr) * 2008-10-08 2010-04-15 Agency For Science, Technology And Research Appareil pour la culture de cellules ancrage-dépendantes

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11613727B2 (en) 2010-10-08 2023-03-28 Terumo Bct, Inc. Configurable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11773363B2 (en) 2010-10-08 2023-10-03 Terumo Bct, Inc. Configurable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11746319B2 (en) 2010-10-08 2023-09-05 Terumo Bct, Inc. Customizable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11708554B2 (en) 2013-11-16 2023-07-25 Terumo Bct, Inc. Expanding cells in a bioreactor
US11667876B2 (en) 2013-11-16 2023-06-06 Terumo Bct, Inc. Expanding cells in a bioreactor
US11795432B2 (en) 2014-03-25 2023-10-24 Terumo Bct, Inc. Passive replacement of media
US11667881B2 (en) 2014-09-26 2023-06-06 Terumo Bct, Inc. Scheduled feed
US11608486B2 (en) 2015-07-02 2023-03-21 Terumo Bct, Inc. Cell growth with mechanical stimuli
US11965175B2 (en) 2016-05-25 2024-04-23 Terumo Bct, Inc. Cell expansion
US11685883B2 (en) 2016-06-07 2023-06-27 Terumo Bct, Inc. Methods and systems for coating a cell growth surface
US11634677B2 (en) 2016-06-07 2023-04-25 Terumo Bct, Inc. Coating a bioreactor in a cell expansion system
US11702634B2 (en) 2017-03-31 2023-07-18 Terumo Bct, Inc. Expanding cells in a bioreactor
US11629332B2 (en) 2017-03-31 2023-04-18 Terumo Bct, Inc. Cell expansion
US11624046B2 (en) 2017-03-31 2023-04-11 Terumo Bct, Inc. Cell expansion
CN108144559A (zh) * 2018-02-24 2018-06-12 清华大学 一种基于核径迹膜的催化膜反应器及其制备方法
US11999929B2 (en) 2020-04-10 2024-06-04 Terumo Bct, Inc. Methods and systems for coating a cell growth surface

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Publication number Publication date
DE102013203306B4 (de) 2015-10-22
DE102013203306A1 (de) 2014-08-28

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