WO1998031783A1 - Systeme de culture de cellules - Google Patents

Systeme de culture de cellules Download PDF

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Publication number
WO1998031783A1
WO1998031783A1 PCT/GB1998/000116 GB9800116W WO9831783A1 WO 1998031783 A1 WO1998031783 A1 WO 1998031783A1 GB 9800116 W GB9800116 W GB 9800116W WO 9831783 A1 WO9831783 A1 WO 9831783A1
Authority
WO
WIPO (PCT)
Prior art keywords
frame
cells
culture substrate
cell culture
substrate
Prior art date
Application number
PCT/GB1998/000116
Other languages
English (en)
Inventor
Patrick Lewis Blott
Dawn Poole
Original Assignee
Smith & Nephew, Plc
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 Smith & Nephew, Plc filed Critical Smith & Nephew, Plc
Priority to AU56706/98A priority Critical patent/AU732565B2/en
Priority to JP53393398A priority patent/JP2001509023A/ja
Priority to CA002277815A priority patent/CA2277815A1/fr
Priority to EP98900896A priority patent/EP0954561A1/fr
Publication of WO1998031783A1 publication Critical patent/WO1998031783A1/fr

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
    • C12M23/02—Form or structure of the vessel
    • C12M23/04—Flat or tray type, drawers
    • 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
    • C12M23/10—Petri dish
    • 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
    • C12M23/20—Material Coatings
    • 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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/06—Plates; Walls; Drawers; Multilayer plates

Definitions

  • the present invention relates to substrates and vessels used for cell-culture techniques.
  • substrates on which cells may be grown for subsequent transfer to a patient requiring a layer of cells are particularly preferred.
  • W091/13638 describes a wound dressing which comprises a layer of a polymeric material having a layer of cultured mammalian cells anchored to one surface.
  • a wound dressing which comprises a layer of a polymeric material having a layer of cultured mammalian cells anchored to one surface.
  • the cells are preferably epithelial cells which may be cultured from autologous skin cells taken by biopsy or may be from other sources.
  • the cells are cultured and grown in the presence of the polymer substrate to which they attach. When the substrate is applied to a prepared site on the patient the cells transfer from the substrate to the patient and continue to multiply to form a layer of epidermal cells.
  • the dressings described are made by culturing the cells in contact with the surface of the polymeric dressing substrate, removing the substrate form the culture medium, and then placing the dressing on the patient such that the cells contact the wound- bed on the patient.
  • An object of the present invention is to provide an improved form of cell culture substrate device which is convenient to use in the culture and anchorage of cells to the substrate and subsequent transfer of cells to the patient. It is a further object to provide a cell culture system which also achieves the above-mentioned objective.
  • a cell culture substrate device comprises a frame formed from a material which has no or low toxicity to cells and a cell culture substrate which is not toxic to cells and having at least one surface to which cells may attach, said frame being adapted to releasably secure said cell culture substrate in a suitable configuration for the attachment of cells to said at least one surface.
  • the frame is preferably of a suitable shape and size to fit into a cell-culture vessel. Preferred forms are generally square, rectangular or round although other shapes may be equally suitable.
  • the frame is adapted to secure a cell culture substrate in a suitable configuration for the attachment of cells to a surface thereof.
  • the cell culture substrate preferably comprises a polymeric film which has a surface suitable for the attachment of cells.
  • the film may be either hydrophilic or hydrophobia
  • the substrate may comprise more than one layer of material.
  • Suitable cell culture substrate materials include those mentioned in W091/13638, i.e. polyesters, polypropylene, blends or copolymers of vinyl acetate, e.g. ethylene - vinyl acetate copolymer, polyvinylidene chloride, polystyrene, polybutadienes, polyethylene, ionomers, e.g. SURLYN (trade mark), or copolymers and blends containing these compounds.
  • hydrophilic polymers such as hydrophilic polyurethanes may form the culture substrate.
  • the culture substrate has a first surface to which cultured mammalian cells may attach. This surface may be treated to modify its properties such as by coating with another material such as a collagen-based material, or by other means, e.g. corona-discharge or plasma treatment.
  • the substrate material is preferably a film of polymeric material.
  • the substrate material is relatively conformable to body contours.
  • the substrate may be continuous or may be apertured.
  • the cell culture substrate material is preferably suitable for application to a wound to serve as a wound dressing. Therefore the cell culture substrate is preferably conformable and elastomeric so as to conform to the contours of the wound to which it is applied.
  • the material may be breathable.
  • the cell culture substrate has epithelial cells attached to a surface thereof, it may be applied to a wound such that the cells are in contact with the wound to effect transfer of cells from the substrate to the wound to promote re- epithelialisation of the wound.
  • the substrate comprises a planar material such as a polymeric film
  • the frame is preferably adapted to secure the substrate in a substantially flat configuration. In this way pooling of cells on the substrate may be minimised so that cells can be relatively evenly distributed upon the surface to which they attach.
  • the frame preferably comprises a planar portion which engages at least a part of said cell culture substrate to releasably secure the substrate in a flat configuration in a plane which is parallel to said planar portion.
  • the planar portion may comprise an open framework or a continuous layer of material.
  • the planar portion comprises an open frame comprising a peripheral i.e. enclosed band, frame to which the cell culture substrate may be attached.
  • the cell culture substrate may be releasably secured to the frame e.g. by adhesive bonding, lamination, heat or radio-frequency welding or mechanical means such as clips.
  • the bond between the frame and the cell culture substrate is preferably liquid-tight.
  • the frame may serve to press against part or all of the substrate to maintain it in a flat configuration against another surface which may be the inner surface of a container or cell culture vessel.
  • the frame may further comprise upstanding walls which may or may not extend continuously around said planar portion.
  • the walls may comprise means for retaining said frame within a container or vessel.
  • the walls may include a lip or recess which may engage the edge of a container such as a petri dish or cell culture vessel.
  • the walls are integral with the other parts of the frame.
  • the frame is made form materials which are non-toxic or not significantly toxic to cells. Suitable materials include polyesters (e.g. polyethylene terephthalate, polyethylene naphthalate), including blends and copolymers thereof, polyolefms, polystyrene, polycarbonate, acrylic materials, polyetherimides, ethylene-vinyl acetate copolymers and blends and copolymers containing any of these or other suitable materials.
  • the frame may be formed by moulding e.g. blow moulding, thermoforming, vacuum or pressure forming or injection moulding.
  • the frame may be formed from a film of polymer by vacuum forming and heating to form the desired shape.
  • a cell culture system comprises a device comprising a frame formed from a material which has no or low toxicity to cells and a cell culture substrate which is not toxic to cells and having at least one surface to which cells may attach, said frame being adapted to releasably secure said cell culture substrate in a suitable configuration for the attachment of cells to said surface and a container wherein said frame is of a shape and size to fit into said container such that said cell culture substrate is held substantially parallel to one internal surface of said container.
  • the culture substrate may further comprise a support layer.
  • the culture substrate may be releasably attached to the support layer over a part or all of one of its surfaces.
  • the surface to which a support layer is attached is opposed to the surface to which cells attach.
  • the culture substrate and support layer are secured in a flat configuration by the frame.
  • the support layer preferably comprises a layer of a polymeric film material which is stiffer than the culture substrate and thus it helps to maintain the culture substrate flat and free of wrinkles.
  • the support layer may comprise a film of a material similar to those which are suitable to form the frame.
  • the support layer is preferably releasably attached over the entire surface of the rest of the culture substrate which is opposed to the surface to which cells attach.
  • the support is preferably attached in such a way that it may be removed from the rest of the culture substrate by peeling when required.
  • the cell culture substrate material comprises a first surface to which cultured mammalian cells may attach and a second opposed surface which is releasably attached over at least the greater part of its second surface to the first surface of a continuous polymeric film which forms the support layer of the culture substrate.
  • the second surface of the cell culture is releasably attached to a portion, preferably central portion, of the support layer.
  • the remaining part of the support layer not in contact with the second opposed surface of the substrate material may be releasably or fixedly attached to the frame.
  • a cell culture substrate device comprising a frame formed from a material which has no or low toxicity to cells and a cell culture substrate which is not toxic to cells and having at least one surface to which cells may attach, wherein a portion of the substrate is releasably attached to the other portion of the substrate, said other portion being fixedly attached to said frame.
  • Fig 1 a perspective view of a device according to the invention
  • FIG 2 a cross sectional view of the device shown in Fig 1 ; placed within a vessel
  • Fig 3 a cross section through a second embodiment.
  • FIG. 4 a perspective view of a device according to a third embodiment placed within a vessel
  • Fig. 5 a cross-section of view of the device according to Fig.4.
  • Fig 1 shows a cell culture device comprising a frame 10 and a cell culture substrate 14.
  • the frame 10 comprises an open peripheral frame 11, which forms a planar portion of the frame, and upstanding walls 13. Walls 13 and peripheral frame 11 are formed integrally by vacuum forming a sheet of 200 ⁇ m thick polyethylene terephthalate film. The walls 13 have a lip 12 extending around their upper edges.
  • the cell culture substrate film 14 is releasably secured to the lower surface of peripheral frame 11 by lamination.
  • the cell culture substrate comprises a conformable sheet of ethylene-vinyl acetate film which has been treated on its upper surface by a corona discharge technique to modify its surface properties for optimal cell attachment.
  • the central portion of this upper surface is accessible through the open part of peripheral frame 11.
  • the frame and culture substrate which form the device may be placed within a petri-dish 15 as shown in Fig 2.
  • the frame and culture substrate are preferably sterile and supplied in a bacteria-proof package.
  • the cell culture device To use the cell culture device, it is placed in a petri-dish or other container and then a cell culture medium containing e.g. human epithelial keratinocytes is added.
  • the cells are cultured at 37°C for a number of days during which the cells multiply and attach to the substrate until the required cell density and degree of confluence of the cell layer is achieved.
  • the medium is then removed and the device is removed from the container.
  • the cell culture substrate may then be peeled from the frame and placed, cell side downwards, on a wound to be treated.
  • the culture substrate is attached over its lower surface to a support layer 16 which is a 150 ⁇ m thick piece of PET film.
  • the support provides additional rigidity to the film 14 to help maintain it in a flat and wrinkle-free condition.
  • the cell culture substrate material 14 is peelably attached on its lower surface to support layer 16 over that part of support layer 16 defined by the peripheral frame 11.
  • substrate material 14 is provided with a tab portion 17 towards a corner of substrate material 14. The remaining part of support layer 16 is fixedly attached to frame 11.

Landscapes

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

Abstract

La présente invention concerne un dispositif de substrat de culture de cellules comprenant un cadre et un matériau substrat destiné à la culture des cellules. Le cadre est conçu de manière qu'il permet de fixer de façon libérable et de préférence selon une configuration plate le matériau substrat sur lequel les cellules viendront s'attacher. Le matériau substrat peut également comporter une couche support. Le dispositif peut être placé dans un conteneur, par exemple une boîte de Petri.
PCT/GB1998/000116 1997-01-16 1998-01-14 Systeme de culture de cellules WO1998031783A1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
AU56706/98A AU732565B2 (en) 1997-01-16 1998-01-14 Cell-culture system
JP53393398A JP2001509023A (ja) 1997-01-16 1998-01-14 細胞培養システム
CA002277815A CA2277815A1 (fr) 1997-01-16 1998-01-14 Systeme de culture de cellules
EP98900896A EP0954561A1 (fr) 1997-01-16 1998-01-14 Systeme de culture de cellules

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9700840.3 1997-01-16
GBGB9700840.3A GB9700840D0 (en) 1997-01-16 1997-01-16 Cell-culture system

Publications (1)

Publication Number Publication Date
WO1998031783A1 true WO1998031783A1 (fr) 1998-07-23

Family

ID=10806084

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1998/000116 WO1998031783A1 (fr) 1997-01-16 1998-01-14 Systeme de culture de cellules

Country Status (8)

Country Link
EP (1) EP0954561A1 (fr)
JP (1) JP2001509023A (fr)
CN (1) CN1249778A (fr)
AU (1) AU732565B2 (fr)
CA (1) CA2277815A1 (fr)
GB (1) GB9700840D0 (fr)
WO (1) WO1998031783A1 (fr)
ZA (1) ZA98329B (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009114138A2 (fr) * 2008-03-14 2009-09-17 Corning Incorporated Dispositif de culture cellulaire et procédés de fabrication et d’utilisation du dispositif de culture cellulaire
DE102013101456A1 (de) * 2013-02-14 2014-08-14 Angelika Spindler Kulturgefäße zur Aufnahme eines Nährmediums für die Inokolum- oder Sporenproduktion im Rahmen einer in-vitro Massenproduktion von VA-Mykorrhizasporen
US9745546B2 (en) 2011-11-07 2017-08-29 Rapid Micro Biosystems, Inc. Cassette for sterility testing
US10407707B2 (en) 2012-04-16 2019-09-10 Rapid Micro Biosystems, Inc. Cell culturing device

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101245315B (zh) * 2008-03-13 2011-04-13 上海交通大学 实验室贴壁细胞薄膜培养装置及方法
JP5942387B2 (ja) * 2011-11-08 2016-06-29 大日本印刷株式会社 細胞培養容器及び培養細胞回収方法
EP3431582A1 (fr) * 2017-07-18 2019-01-23 Koninklijke Philips N.V. Matériaux de culture de cellules
KR102181128B1 (ko) * 2019-02-27 2020-11-20 경북대학교 산학협력단 배양 용기에 고정 가능한 세포 배양용 나노섬유 매트 및 이를 배양 용기 내에 고정하는 방법
CN112221543B (zh) * 2019-07-15 2024-05-10 京东方科技集团股份有限公司 细胞片层转移器件、运输装置和转移细胞片层的方法
WO2022202652A1 (fr) * 2021-03-23 2022-09-29 国立大学法人東海国立大学機構 Agent de revêtement pour culture cellulaire, récipient de culture cellulaire et son procédé de production, et procédé de culture cellulaire

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1017751B (de) * 1956-01-19 1957-10-17 Dr Herbert Bucherer Geraet zur Zuechtung von Mikroorganismen
EP0239697A2 (fr) * 1986-03-20 1987-10-07 Costar Corporation Appareil pour la culture de tissu in vitro
US4715911A (en) * 1985-03-25 1987-12-29 A/S Nunc Method for the production of a plastic container
US4748124A (en) * 1984-10-30 1988-05-31 E. I. Du Pont De Nemours And Company Compartmentalized cell-culture device and method
EP0307048A2 (fr) * 1987-09-11 1989-03-15 Joseph Guy Cremonese Flacon pour la culture de cellules muni d'une membrane
US5190878A (en) * 1989-07-14 1993-03-02 Minuth Wilhelm Apparatus for cultivating cells
US5264344A (en) * 1989-03-04 1993-11-23 University Of Leicester Method for culturing of cells on a membrane
GB2268187A (en) * 1992-07-01 1994-01-05 Univ Hull Cell culture vessels
JPH0923876A (ja) * 1995-07-11 1997-01-28 Kao Corp 細胞培養支持体の製造法

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1017751B (de) * 1956-01-19 1957-10-17 Dr Herbert Bucherer Geraet zur Zuechtung von Mikroorganismen
US4748124A (en) * 1984-10-30 1988-05-31 E. I. Du Pont De Nemours And Company Compartmentalized cell-culture device and method
US4715911A (en) * 1985-03-25 1987-12-29 A/S Nunc Method for the production of a plastic container
EP0239697A2 (fr) * 1986-03-20 1987-10-07 Costar Corporation Appareil pour la culture de tissu in vitro
EP0307048A2 (fr) * 1987-09-11 1989-03-15 Joseph Guy Cremonese Flacon pour la culture de cellules muni d'une membrane
US5264344A (en) * 1989-03-04 1993-11-23 University Of Leicester Method for culturing of cells on a membrane
US5190878A (en) * 1989-07-14 1993-03-02 Minuth Wilhelm Apparatus for cultivating cells
GB2268187A (en) * 1992-07-01 1994-01-05 Univ Hull Cell culture vessels
JPH0923876A (ja) * 1995-07-11 1997-01-28 Kao Corp 細胞培養支持体の製造法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
PATENT ABSTRACTS OF JAPAN vol. 097, no. 005 30 May 1997 (1997-05-30) *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009114138A2 (fr) * 2008-03-14 2009-09-17 Corning Incorporated Dispositif de culture cellulaire et procédés de fabrication et d’utilisation du dispositif de culture cellulaire
WO2009114138A3 (fr) * 2008-03-14 2010-02-04 Corning Incorporated Dispositif de culture cellulaire et procédés de fabrication et d’utilisation du dispositif de culture cellulaire
US9745546B2 (en) 2011-11-07 2017-08-29 Rapid Micro Biosystems, Inc. Cassette for sterility testing
US10801004B2 (en) 2011-11-07 2020-10-13 Rapid Micro Biosystems, Inc. Cassette for sterility testing
US11788046B2 (en) 2011-11-07 2023-10-17 Rapid Micro Biosystems, Inc. Cassette for sterility testing
US10407707B2 (en) 2012-04-16 2019-09-10 Rapid Micro Biosystems, Inc. Cell culturing device
US11643677B2 (en) 2012-04-16 2023-05-09 Rapid Micro Biosystems, Inc. Cell culturing device
DE102013101456A1 (de) * 2013-02-14 2014-08-14 Angelika Spindler Kulturgefäße zur Aufnahme eines Nährmediums für die Inokolum- oder Sporenproduktion im Rahmen einer in-vitro Massenproduktion von VA-Mykorrhizasporen
DE102013101456B4 (de) * 2013-02-14 2014-12-11 Angelika Spindler Kulturgefäße zur Aufnahme eines Nährmediums für die Inokolum- oder Sporenproduktion im Rahmen einer in-vitro Massenproduktion von VA-Mykorrhizasporen

Also Published As

Publication number Publication date
JP2001509023A (ja) 2001-07-10
AU732565B2 (en) 2001-04-26
CA2277815A1 (fr) 1998-07-23
ZA98329B (en) 1998-07-16
EP0954561A1 (fr) 1999-11-10
CN1249778A (zh) 2000-04-05
GB9700840D0 (en) 1997-03-05
AU5670698A (en) 1998-08-07

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