EP2257376A1 - Plaque de culture à puits multiples destinée à des cultures tridimensionnelles - Google Patents

Plaque de culture à puits multiples destinée à des cultures tridimensionnelles

Info

Publication number
EP2257376A1
EP2257376A1 EP09712690A EP09712690A EP2257376A1 EP 2257376 A1 EP2257376 A1 EP 2257376A1 EP 09712690 A EP09712690 A EP 09712690A EP 09712690 A EP09712690 A EP 09712690A EP 2257376 A1 EP2257376 A1 EP 2257376A1
Authority
EP
European Patent Office
Prior art keywords
bottom part
multiwell plate
wall
cavity
multiwell
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.)
Withdrawn
Application number
EP09712690A
Other languages
German (de)
English (en)
Inventor
Daniela Marino
Michael Detmar
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.)
Eidgenoessische Technische Hochschule Zurich ETHZ
Original Assignee
Eidgenoessische Technische Hochschule Zurich ETHZ
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 Eidgenoessische Technische Hochschule Zurich ETHZ filed Critical Eidgenoessische Technische Hochschule Zurich ETHZ
Priority to EP09712690A priority Critical patent/EP2257376A1/fr
Publication of EP2257376A1 publication Critical patent/EP2257376A1/fr
Withdrawn legal-status Critical Current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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/22—Transparent or translucent parts
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
    • B01L3/50855—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates using modular assemblies of strips or of individual wells
    • 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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/14—Process control and prevention of errors
    • B01L2200/141—Preventing contamination, tampering
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/041—Connecting closures to device or container
    • B01L2300/045—Connecting closures to device or container whereby the whole cover is slidable
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0829—Multi-well plates; Microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0848—Specific forms of parts of containers
    • B01L2300/0851—Bottom walls
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/505—Containers for the purpose of retaining a material to be analysed, e.g. test tubes flexible containers not provided for above

Definitions

  • the invention relates to multiwell culture plates for growing cell or tissue cultures in vitro, and a method for transferring a cell or tissue culture from a multiwell plate, according to the preambles of the independent claims.
  • multiwell culture plates consisting of a plate body having a multitudes of wells.
  • Industry stan- dard multiwell plates are laid out with 96 wells, arranged in an 8 x 12 matrix.
  • Other variants of multiwell plates are available, e.g. with 6, 12, 24, 48, 96, 384, or 1536 wells.
  • the size of the wells, the distance between the rows and columns, as well as the height, length and width of said multiwell plates are standardized.
  • a large number of auxiliary equipment is available for such standard size multiwell plates, including devices that load and unload precise volumes of liquid in multiples of 8, 12, or 96 wells at a time.
  • Most multiwell plates for cell cultures are consumable goods intended for single-use, manufactured of suitable plastic material such as polystyrene.
  • WO 86/07606 discloses a multiwell plate with wells that contain an opening in the bottom, and a filter membrane arranged at that opening. Such a multiwell plate is intended to gain the liquid medium of a number of bacteria or cell cultures in parallel.
  • WO 98/35013 shows a multiwell plate with wells that are di- vided into two chambers by a semi-permeable membrane placed in the middle of the well.
  • the known multiwell culture plates essentially comprise a rigid plate frame structure with a multitude of cavities, regularly arranged on the plate body, each cavity having an opening on top of the plate. A cover can be placed on the plate in order to protect the cultures from contamination.
  • the multiwell plates have to be completely sterile, which is achieved for example by treating the multiwell plates in their sealed packages with gamma radiation.
  • Another problem is the lack of reproducibility of such manual transfer steps, particu- larly its influence on the cell culture. As a consequence the culturing of three- dimensional cell samples cannot be standardized to the same degree as it is possible for cultures in liquid media.
  • An object of the invention is to provide multiwell culture plates for growing cell or tis- sue cultures in vitro that do not have the above-mentioned and other disadvantages, and particularly do allow the transfer of cell cultures from specific wells of the multiwell plate with reduced mechanical stress or physical deformation.
  • Another object of the invention is to provide a method for transferring a cell or tissue culture from a multiwell plate, in which the cultures are subject to less mechanical stress or physical deformation.
  • a multiwell culture plate according to the invention and the method according to the invention should give the possibility to automatize such transfers, for example by using laboratory robots.
  • Another object of the invention is an improved reproducibility of the transfer step, particularly its influence on the cell culture.
  • the cavities are each formed by a rigid wall and a preferably flat bottom part, said bottom part being hermetically bonded to the cavity wall, while at the same time being severable or sectionable from the wall along its rim.
  • the bottom part of the cavity is detached from the cavity wall with a suitable detaching device that allows to sever, to section or to punch the bottom part along its rim, or to break the bonding between bottom and wall, e.g. by applying pressure, or to break a predetermined breaking point along the rim of the bottom part.
  • the three-dimensional culture remains on the detached bottom part, while the multiwell plate is lifted away, perpendicular to the bot- torn part.
  • An important characteristics of the invention is that the bottom part of each individual cavity consists of a thin, transparent, non-porous, cell culture compatible bottom that can be individually removed from the multiwell plate for example by a cutting device, together with the cell or tissue culture 4 that had been cultured on it.
  • multiwell plates according to the invention are especially advantageous for three-dimensional cell cultures, organotypic cultures, tissue engineering and regeneration, scaffold generation and synthesis, three-dimensional cultures of organs or parts of organs, e.g. chicken spinal cord in a agarose matrix, and im- aging of three-dimensional biological structures.
  • the bottom part has to be waterproof, non-porous, and cell culture compatible.
  • the bottom part is a semi-flexible sheet or membrane. Even more preferably, the bottom part is a thin membrane, most preferably five to five hundred micrometers thin.
  • Suitable materials for the bottom part are for example polycarbonate, polyester, or polytetrafluoroethylene. Espe- cially preferred materials are polystyrene and polyethylene.
  • the membrane must be compatible with routine tissue embedding and sectioning procedures, such as embedding in paraffin and sectioning with a cryomicrotome or a microtome for paraffin sections.
  • the bottom part of the cavity may be attached to the cavity wall for example by adhesive, or thermal or ultrasonic welding.
  • the bottom part is realized as a semi- flexible membrane, it is possible to attach one sheet of membrane, subsequently forming the bottom parts of several cavities. It is also possible to attach the membrane with a clamping mechanism as shown for example in WO 86/07606.
  • the bottom part is translucent or transparent, to allow micros- copy of the culture sample while illuminating the sample through the bottom part.
  • the multiwell plate comprises a drawer element arranged on the plate frame structure close to the bottom parts of the cavities.
  • the drawer element During normal handling, e.g. transporting or storing the multiwell plate, the drawer element remains in an inserted position, protecting the bottom part of the cavities from damages.
  • the drawer element For transferring a culture from a specific cavity, the drawer element is retracted as far as necessary to free the specific cavity bottom. After removing the culture, the drawer is closed again, protecting the remaining cavity bottom parts.
  • two or more drawer elements may be used, each giving e.g. access to one or more rows or columns of the multiwell plate according to the invention.
  • a method for transferring a cell or tissue culture from a multiwell plate with a multitude of cavities, having their only opening oriented towards a first side of the multiwell plate basically comprises the steps of (a) detaching the bottom part from the wall of a specific cavity, thereby forming a new opening in the cavity opposite to the first side of the plate; and (b) removing the culture through said new opening together with the detached cavity bottom part.
  • the bottom part is detached by severing, sectioning, or punching the bottom part along its rim, or by breaking the bonding between the bottom part and the wall, or by breaking a predetermined breaking point along the rim of the bottom part.
  • the culture is not subject to mechanical stress and is not deformed, the negative influence of the transfer step is much less that with the methods known from the prior art.
  • the method can be carried out in a standardized, reproducible manner, thereby decreasing irreproducible errors in the experiment. Furthermore the method can be easily carried out by laboratory robots, and is highly automizable.
  • Figure 1 schematically shows a preferred embodiment of a multiwell plate according to the invention, having two drawer elements, (a) with fully inserted drawer elements, and (b) with one drawer element partially pulled out, and one three-dimensional culture removed.
  • Figure 2 discloses (a) a device for severing or sectioning the bottom part of a well of a multiwell plate according to the invention along its rim, and (b) the application of said device
  • Figure 3 shows yet another embodiment of a multiwell plate according to the invention, with a reusable plate structure and modular, replaceable cavity inserts, (a) in a side-view, and (b) in a transverse sectional view.
  • Figure 4 shows a transverse sectional view of yet another embodiment, with modular cavity inserts and drawers.
  • Figure 5 shows a possible application of a multiwell plate according to the invention.
  • FIG. l(a) An embodiment of a multiwell plate 1 according to the invention with two drawer elements 3, 3', is schematically shown in Figure l(a) with fully inserted drawer elements 3, 3', and in Figure l(b) with one drawer element 3' partially pulled out, and one three- dimensional culture 4 removed from the cavity 2'.
  • the shown multiwell plate 1 comprises a plate frame structure 11 and six cavities 2.
  • the cavities 2 comprise a cylindrical wall 21 and a bottom part 22, said bottom part being severable or sectionable along its rim 23.
  • the bottom part is a semi-flexible membrane, which is stable enough to carry the weight of heavy tissue scaffolds without being damaged, and thin enough to be easily detached from the cavity wall when necessary.
  • the multiwell plate 1 also comprises a cover 12, shown above the plate 1.
  • Said cover 12 has no effect on the function of the invention itself.
  • Two drawer elements 3, 3' are arranged in the plate frame structure 11, below the bottom parts 22.
  • the drawer elements 3, 3' comprise a handle 31 and a drawer base 32, and are movable on tracks along the long axis of the multiwell plate 1.
  • the drawer base 32 is advantageously positioned close to the bottom part membrane 22, but preferably has no direct contact with the membrane, to avoid unnecessary mechanical movements of the culture when the drawer is opened and closed.
  • a special device 5 as shown in Figure 2 may be used.
  • Such a device 8 essentially consists of a handle 52 and a circularly ar- ranged blade or wire 51.
  • the multiwell plate 1 is placed on a suitable flat surface, and the drawer element 3' is partially pulled out.
  • the device 5 is used to sever or cut the bottom part membrane 22.
  • the blade or wire 52 of the device 5 has a diameter that is slightly smaller than the inner diameter of the cylindrical cavity wall 21.
  • a third step after detaching the bottom part 22 from the cavity 2, the multi- well plate 1 is lifted, and the three-dimensional culture 4 remains on the surface, together with the detached bottom part membrane 22.
  • FIG. 3(a) Another embodiment of a multiwell plate 1 according to the invention is schematically shown in Figure 3(a) in a side-view, and Figure 3(b) in a transverse sectional view, hi this embodiment of the invention the plate frame structure 11 may be reused, while only the replaceable cavities 2 are single-use.
  • the multiwell plate 1 comprises a plate frame structure 11 with six circular support structures 13 for cavity inserts 24. Said single-use cavity inserts form the cavities 2" of the multiwell plate, comprising a cylindrical wall 21 and bottom part 22.
  • the bottom part 22 is again severable or sectionable from the cavity 2 along its rim 23.
  • the cavity insert 24 is placed on the support structure 13.
  • the shown embodiment comprises no drawers, but is permanently closed by a plate base 15.
  • the insert 24 with cavity 2" holding the culture is removed from the multiwell plate 1 and placed on surface.
  • the bottom part 22 is then detached from the cavity 2" in the same manner as for the embodiment shown in Figure 1.
  • the cavities are modular as in the embodiment of Figure 3, and the plate structure 1 comprises drawers 3, 3', as in the embodiment of Figure 1.
  • FIG. 5 Another advantage of modular cavity inserts is that each cavity may be equipped with its own cover.
  • the application of a multiwell plate 1 according to the invention is described in Figure 5.
  • the cover 12 is removed from the plate 1 , and cell samples 6 are placed in the cavities 2, together with a gel or a scaffold as the three-dimensional medium.
  • the cover 12 is closed, and the cells are cultured.
  • the cover 12 is removed and the drawer 3 is partially retracted.
  • the culture sample 4 is then removed, together with the bottom part 22, and further processed (fourth step), e.g. placed in an embedding mould.
  • the cover 12 is closed again and the culturing of the remaining cell cultures is continued.

Landscapes

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

Abstract

Dans une plaque à puits multiples 1, chacune des cavités de la multitude de cavités 2 est constituée par une paroi rigide 21 et une partie inférieure plate 22, ladite partie inférieure étant collée de manière hermétique à la paroi de cavité 21, tout en pouvant être détachée de façon individuelle de la paroi 21 le long de son rebord 23. Pour transférer une culture tridimensionnelle 4 à un deuxième dispositif, la partie inférieure 22 de la cavité 2 est détachée de façon individuelle de la paroi de cavité avec un dispositif approprié 5, qui permet de détacher la partie inférieure le long de son rebord, par exemple en appliquant une pression. La culture tridimensionnelle 4 reste sur la partie inférieure détachée, tandis que la plaque à puits multiple avec la paroi de cavité est soulevée. Etant donné que la culture 4 elle-même n'est pas contrainte de manière mécanique, elle n’est pas endommagée.
EP09712690A 2008-02-20 2009-02-04 Plaque de culture à puits multiples destinée à des cultures tridimensionnelles Withdrawn EP2257376A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP09712690A EP2257376A1 (fr) 2008-02-20 2009-02-04 Plaque de culture à puits multiples destinée à des cultures tridimensionnelles

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
EP08003064 2008-02-20
EP09712690A EP2257376A1 (fr) 2008-02-20 2009-02-04 Plaque de culture à puits multiples destinée à des cultures tridimensionnelles
PCT/EP2009/000724 WO2009103416A1 (fr) 2008-02-20 2009-02-04 Plaque de culture à puits multiples destinée à des cultures tridimensionnelles

Publications (1)

Publication Number Publication Date
EP2257376A1 true EP2257376A1 (fr) 2010-12-08

Family

ID=39637600

Family Applications (1)

Application Number Title Priority Date Filing Date
EP09712690A Withdrawn EP2257376A1 (fr) 2008-02-20 2009-02-04 Plaque de culture à puits multiples destinée à des cultures tridimensionnelles

Country Status (2)

Country Link
EP (1) EP2257376A1 (fr)
WO (1) WO2009103416A1 (fr)

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB201012490D0 (en) 2010-07-26 2010-09-08 Randox Lab Ltd Sealed well assembly, cartridge thereof, and apparatus and method for opening a sealed well assembly
WO2012144983A1 (fr) * 2011-04-18 2012-10-26 Empire Technology Development Llc Cartouche de culture cellulaire rotative et ses méthodes d'utilisation
CN103146572B (zh) * 2011-12-07 2016-01-06 清华大学 一种实现细胞集落均质性生长的装置和方法
ITUB20151992A1 (it) 2015-07-08 2017-01-08 Ivtech S R L Piastra multi-pozzetto per colture cellulari
WO2017059273A1 (fr) * 2015-10-01 2017-04-06 Berkeley Lights, Inc. Incubateur à plaque à cavités
JP6968169B2 (ja) 2016-12-01 2021-11-17 バークレー ライツ，インコーポレイテッド ウェルプレートインキュベーター
IT201900005820A1 (it) * 2019-04-15 2020-10-15 Univ Bologna Alma Mater Studiorum Dispositivo per colture cellulari
CN112358966B (zh) * 2020-10-23 2022-03-22 上海交通大学 一种滑动式微流控器官芯片及其制备方法

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US3883398A (en) * 1973-05-07 1975-05-13 Bellco Glass Inc Microculture slide chamber
DE2902026C3 (de) * 1979-01-19 1981-10-29 Peters, J. Hinrich, Dr., 5064 Rösrath Biologisches Gefäß
US7371563B2 (en) * 2000-11-08 2008-05-13 Surface Logix, Inc. Peelable and resealable devices for biochemical assays
DE60122854T2 (de) * 2001-02-20 2007-03-08 F. Hoffmann-La Roche Ag Lineare Küvettenmatrix, damit gebaute zweidimensionale Küvettenmatrix und solche zweidimensionale Küvettenmatrizen umfassendes System
US6716350B2 (en) * 2002-05-03 2004-04-06 Millipore Corporation Microplate protective tray undercover
EP1477792B1 (fr) * 2003-05-13 2006-10-25 Becton, Dickinson and Company Procédé et dispositif pour purifier et dessaler des échantillons biologiques

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Title
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Also Published As

Publication number Publication date
WO2009103416A1 (fr) 2009-08-27

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