EP3724315A1 - Agencement de microbioréacteur - Google Patents

Agencement de microbioréacteur

Info

Publication number
EP3724315A1
EP3724315A1 EP17828878.3A EP17828878A EP3724315A1 EP 3724315 A1 EP3724315 A1 EP 3724315A1 EP 17828878 A EP17828878 A EP 17828878A EP 3724315 A1 EP3724315 A1 EP 3724315A1
Authority
EP
European Patent Office
Prior art keywords
microbioreactor
arrangement
insert
unit
microtiter plate
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.)
Pending
Application number
EP17828878.3A
Other languages
German (de)
English (en)
Inventor
Andreas Schober
Frank Weise
Jörg HAMPL
Gregor Schlingloff
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.)
Technische Universitaet Ilmenau
Original Assignee
Technische Universitaet Ilmenau
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 Technische Universitaet Ilmenau filed Critical Technische Universitaet Ilmenau
Publication of EP3724315A1 publication Critical patent/EP3724315A1/fr
Pending legal-status Critical Current

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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/02—Membranes; Filters
    • C12M25/04—Membranes; Filters in combination with well or multiwell plates, i.e. culture inserts
    • 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
    • C12M23/12—Well or multiwell plates
    • 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
    • C12M23/22—Transparent or translucent parts
    • 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
    • C12M23/38—Caps; Covers; Plugs; Pouring means
    • 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
    • C12M23/40—Manifolds; Distribution pieces
    • 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
    • C12M25/14—Scaffolds; Matrices
    • 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
    • C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
    • C12M37/04—Seals

Definitions

  • the invention relates to a microbioreactor arrangement with a plurality of microbioreactors.
  • the assembly comprises a microtiter plate with numerous wells in a predetermined grid arrangement.
  • Microtiter plates are basically known in their design and are used for the investigation of biological substances, for example, the absorption measurement in photometers or screening studies numerous Abwand lungs a base substance.
  • Microtiter plates usually have a rectangular basic shape and consist of plastic or glass.
  • a microtiter plate has numerous cells isolated from each other, so-called wells, which are arranged in a predetermined grid. The exact dimensions are chosen regularly according to an ANSI standard.
  • the wells are available in various shapes, for example with flat bottom or U-shaped tapered bottom in each well. In any case, the bottom surface of the microtiter plate is 29los sen and the top open, so that the cavity of each well is accessible from the top.
  • a microtiter plate with a transparent bottom plate for light and a two open to opposite surfaces cavity plate with matrix-like arrangement of the cavities or wells is known.
  • Soil and cavity plate are insoluble and liquid-tight miteinan connected.
  • the bottom plate has a flat, structurally loose surface and carries at its the wells facing first surface of at least two layers with different index of refraction existing layer system.
  • an insert for a Mikroti terplatte consisting of a support structure, in which at least one recess is introduced, wherein the upper diameter of the recess is selected so that it can be inserted into a recess of the microtiter plate ,
  • the bottom of the well has at least one downwardly formed microcavity.
  • the insert is at least partially provided with pores.
  • WO 2011/035937 A1 describes a microstructured shaped body comprising a film which is divided into undeformed areas and thinned drawing areas. At least in some of the thinned stretching areas, microstructures are formed, wherein pores are formed at least in one of the thinner stretched areas and at least some of the undeformed areas are impermeable.
  • a microstructured ter molded article which has a film-like base body, which comprises a first film layer and a second film layer underneath, wherein the second film layer has recesses with a diameter of less than 2 mm, by deformed areas of the first film layer are formed, are formed by which cavities out. At least some of the deformed regions of the first film layer have pores. The areas of the film-like body are outside of the recesses impenetrable.
  • DE 10 2010 037 968 A1 describes a structure for simulating a sinusoid, which can be inserted into a microtiter plate. The structure comprises a plurality of superimposed layers of a porous material, wherein between the layers in each case a gap is formed. The interstices are connected by channels formed in the layers for forwarding a fluid.
  • an object of the present invention is to provide a micro bioreactor system based on microtiter plates, which allows a parallel supply of a plurality of micro bioreactors, which are formed in the individual wells.
  • the microbioreactor arrangement implements a plurality of microbioreactors and for this purpose comprises a microtiter plate with numerous wells in a predetermined, preferably standardized grid arrangement.
  • the microtiter plate has in a conventional manner a closed bottom surface, which preferably allows optical observation, that lets light rays happen.
  • the individual wells are open at the top of the microtiter plate, so that from there inserts of an insert unit, which is arranged on the upper side of the microtiter plate, can be used.
  • the numerous inserts have the same Rasteranord tion, so that each insert engages in a well.
  • the insert divides the well into at least two regions, in particular a cell cultivation region and a
  • an activation unit is part of the micro bioreactor arrangement.
  • the activation unit is on the
  • Inserted unit and has numerous pumps that are connected to the areas formed by the inserts, namely via supply channels, which allow a fluid or nutrient transport between the two areas.
  • microtiter plate individual cell culturing areas are observable through the underside of the microtiter plate and can be used simultaneously through the upper opening also in automatic pipetting systems and in standard analytical equipment (e.g., fluorescence microplate readers). It is thus the complete compatibility with standardized microtiter plates achieved, incl. all handling and analysis options.
  • each of the inserts of the insert unit has a microstructured Formkör which is arranged parallel to the bottom surface of the microtiter plate and a settlement framework for cell cultures willing- provides.
  • WO 2011/035938 Al be formed.
  • a modified embodiment is characterized in that at least one, preferably several of the inserts have at least two microstructured shaped bodies which are positioned parallel to one another and axially spaced from one another.
  • Frame elements or the like is covered. This allows the optical observation of the substances or cell cultures on the moldings through the bottom surface, what preference, conventional inverse microscopes can be used.
  • the microbioreactor arrangement according to the invention can therefore be clamped in existing observation arrangements such as a conventional microtiter plate in order to perform the observations.
  • a distribution channel system is formed in the activation unit, via which the plurality of pumps are supplied with compressed air to perform the pumping operation.
  • all pumps are driven in parallel, so that in each well diesel ben flow conditions prevail.
  • the pumps transport a nutrient solution to supply the cell cultures that are located on the moldings of the inserts.
  • the pumps can also be driven individually or in groups, for which a targeted control, for example by switching valves, which may be an integral part of the activation unit, is possible.
  • An advantageous embodiment is characterized in that the insert unit comprises a sealing structure.
  • the cell culture open area is cylindrically shaped and coaxially positioned in the associated well.
  • the cover unit is provided above the open areas with a central opening through which substances can be supplied by means of a pipette.
  • the closed area can also be arranged cylindrically and coaxially or be inserted in the open area in the form of tubes or capillaries.
  • the two areas are preferably formed on the sealing structure, inserted into this or formed integrally with the sealing structure.
  • the sealing structure extends substantially in the plane of the microtiter plate and lies flat on this surface.
  • the insert unit comprises a supply channel system, which with each of their
  • Inserts is coupled to allow the supply and removal of a nutrient solution to the inserts.
  • the insert unit can be composed of several sub-layers to form the required cavities.
  • the micro bioreactor assembly further comprises a cover plate which is mounted on the activation unit to close the Mikrobioreak gate assembly upwards.
  • the cover plate can in this case be removed if necessary, for example to make the central openings in the inserts accessible.
  • Fig. 1 is an exploded perspective view of a first
  • Fig. 2 is a plan view of the assembled
  • Fig. 3 is a sectional view of the microbioreactor assembly prior to assembly
  • Fig. 4 is a sectional perspective detail view of a single well with an insert.
  • FIG. 1 shows a microbioreactor arrangement 01 according to the invention in a perspective exploded view.
  • the microbioreactor assembly 01 has a plurality of main groups, namely a microtiter plate 02, a patch unit 03 placed on top and a unit above the insert activation unit 04.
  • a separate cover plate 05 is provided which covers the activation unit at the top.
  • the main groups are stacked so that the microtiter plate 02 acts as a lower support.
  • the main groups are fixed after assembly, but if necessary, releasably connected to each other, for example via Rastele elements 07.
  • a therapiessorg circuit 08 is provided, via which, for example, compressed air and a nutrient solution can be supplied. Other connections (not shown) can be seen easily when needed on the insert unit.
  • Fig. 2 shows the microbioreactor assembly 01 in a plan view, wherein the cover plate 05 is made of transparent material, so that the underlying individual wells 09 of the microtiter plate 02 can be seen.
  • the wells or cups 09 are arranged in the example shown in a 6x4 matrix.
  • other microtiter plates can be used, the other main groups are then accordingly adapt.
  • Fig. 3 shows the microbioreactor assembly 01 again in a cutaway exploded view. It can be seen here that the main groups are stacked on top of each other and the matrix arrangement of the microtiter plate 02 in the overlying layers is repeated. In this way, numerous micro bioreactors are formed, which are all identical in their construction but can be populated with different cell cultures.
  • Fig. 4 shows in a perspective sectional view of such a microbioreactor, which is composed of sections of the aforementioned genann th main groups as follows:
  • an insert 11 which is part of the insert unit 03.
  • the insert 11 has a cylindrical cell culturing area 12 into which biological substances can be introduced and cultured.
  • a microstructured shaped body 13 At the bottom of the cell cultivation area 12 is a microstructured shaped body 13, on which cell cultures can be settled.
  • a bottom surface 14 of the microtiter plate 02 is transparent at least below the microstructured shaped body 13 to an optical To allow observation of the settled cell cultures.
  • the wall of the insert 11 opens into a sealing structure 16 which seals the open cell cultivation area 12 against a closed area 17.
  • the closed portion 17 extends in the dargestell th embodiment coaxially between the inner wall of the well 09 and the outer wall of the insert 11.
  • the insert unit 03 also includes a support frame 18, which stabilizes the gasket structure 16 and further provides a supply channel 19 through which nutrient solution is transportable. To the supply channel 19, a suction capillary 21 is connected, which extends axially in the cell cultivation area 12. Thus, nutrient solution can be circulated between the cell culturing area 12 and the closed area 17 via the supply channel 19.
  • the activation unit 04 is placed above, which is composed in the example shown of a lower plate 22 and an upper plate 23, between which a closed pressure chamber 24 is formed.
  • a membrane 26 is used, which acts on the supply channel 19 and the pressure chamber 24 can be acted upon with compressed air.
  • the membrane 26 acts as a pump which drives the delivery of the nutrient solution in the closed area 17.
  • an upwardly open central opening 27 is provided, which tiv michs Scheme 12 opens into the Zellkul. Via the central opening 27, substances can preferably be used with an automated pipette be added.
  • the central opening 27 can be closed by the cover plate 05, if an addition is not required.

Landscapes

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

Abstract

L'invention concerne un agencement de microbioréacteur (01) comprenant une multitude de microbioréacteurs. L'agencement de microbioréacteur comprend une plaque de microtitrage (02) pourvue d'un grand nombre de puits (09) dans un agencement en réseau prédéfini, une surface de fond (14) fermée et une face supérieure ouverte. En outre, une unité d'insert (03) est disposée sur la face supérieure de la plaque de microtitrage (02) et possède un grand nombre d'inserts (11) dans le même agencement en réseau, un insert (11) se logeant dans un puits (09) respectif et le puits (09) étant divisé en au moins deux zones (12, 17). Enfin, l'agencement de microbioréacteur comprend une unité d'activation (04), qui est placée sur l'unité d'insert (03) et qui possède de nombreuses pompes (26) qui sont raccordées à des canaux d'alimentation (19) respectifs, qui permettent un transport fluidique entre les deux zones (12, 17).
EP17828878.3A 2017-12-15 2017-12-15 Agencement de microbioréacteur Pending EP3724315A1 (fr)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/EP2017/083150 WO2019114996A1 (fr) 2017-12-15 2017-12-15 Agencement de microbioréacteur

Publications (1)

Publication Number Publication Date
EP3724315A1 true EP3724315A1 (fr) 2020-10-21

Family

ID=60957269

Family Applications (1)

Application Number Title Priority Date Filing Date
EP17828878.3A Pending EP3724315A1 (fr) 2017-12-15 2017-12-15 Agencement de microbioréacteur

Country Status (3)

Country Link
US (1) US20210155886A1 (fr)
EP (1) EP3724315A1 (fr)
WO (1) WO2019114996A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP7265645B2 (ja) * 2019-11-14 2023-04-26 富士フイルム株式会社 細胞の保存又は輸送器具及び細胞の輸送方法
EP4015619A1 (fr) * 2020-12-17 2022-06-22 Leica Microsystems CMS GmbH Ensemble microplaque et procédé de transfert d'échantillon au moyen d'un ensemble microplaque

Family Cites Families (18)

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US5863792A (en) * 1997-03-19 1999-01-26 Becton Dickson And Company Culture vessel assembly
US5882922A (en) * 1997-03-19 1999-03-16 Becton Dickinson And Company Culture vessel assembly
US5801055A (en) * 1997-09-10 1998-09-01 Becton Dickinson And Company Multi-well culture dish assembly
DE19806681B4 (de) 1998-02-18 2006-07-27 Carl Zeiss Jena Gmbh Mikrotiterplatte
US7163660B2 (en) * 2000-05-31 2007-01-16 Infineon Technologies Ag Arrangement for taking up liquid analytes
DE10046175A1 (de) * 2000-09-19 2002-03-28 Augustinus Bader Verfahren und Vorrichtung zum Züchten und/oder Behandeln von Zellen
DE10118905A1 (de) * 2001-04-18 2002-10-24 Evotec Ag Verfahren und Vorrichtung zum Kultivieren und/oder Verteilen von Partikeln
JP4631339B2 (ja) * 2004-07-27 2011-02-16 株式会社ニコン 環境制御装置、温度調節装置および環境制御型分析装置
DE202006017853U1 (de) 2006-11-23 2007-01-18 Forschungszentrum Karlsruhe Gmbh Einsatz für eine Mikrotiterplatte
CN101270332B (zh) * 2008-04-30 2012-05-30 广东省人民医院 用于细胞或组织培养和处理的装置和方法
DE102009044113A1 (de) 2009-09-27 2011-04-07 Technische Universität Ilmenau Teilweise perforierter mikrostrukturierter Formkörper und Verfahren zu dessen Herstellung
DE102009044115A1 (de) 2009-09-27 2011-04-07 Technische Universität Ilmenau Mikrostrukturierter Formkörper mit perforierten Teilen und Verfahren zu dessen Herstellung
EP2609189A4 (fr) * 2010-08-24 2015-08-05 Hawley & Hazel Chemical Co Zhongshan Ltd Procédés, dispositifs et utilisations liés aux biofilms
DE102010037968A1 (de) 2010-10-05 2012-04-05 Technische Universität Ilmenau Struktur zur Nachbildung eines Sinusoids und Verfahren zu ihrer Herstellung
CA2804568A1 (fr) * 2011-03-22 2012-09-27 Tokai Hit Co., Ltd. Appareil de culture pour la visualisation microscopique et son procede d'utilisation
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EP3190172A3 (fr) * 2016-01-07 2017-08-30 Vanderbilt University Interconnexions de perfusion multiples des constructions tissulaires techniques et microbioréacteurs, multi-microformulateurs et leurs applications

Also Published As

Publication number Publication date
US20210155886A1 (en) 2021-05-27
WO2019114996A1 (fr) 2019-06-20

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