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WO1993000423A1 - Additif pour milieu de culture a base de chelate de fer - Google Patents

Additif pour milieu de culture a base de chelate de fer Download PDF

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Publication number
WO1993000423A1
WO1993000423A1 PCT/DK1992/000190 DK9200190W WO9300423A1 WO 1993000423 A1 WO1993000423 A1 WO 1993000423A1 DK 9200190 W DK9200190 W DK 9200190W WO 9300423 A1 WO9300423 A1 WO 9300423A1
Authority
WO
WIPO (PCT)
Prior art keywords
citrate
culture medium
fecl
iron
free
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/DK1992/000190
Other languages
English (en)
Inventor
Peter Bernt Suhr-Jessen
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.)
Novo Nordisk AS
Original Assignee
Novo Nordisk AS
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 Novo Nordisk AS filed Critical Novo Nordisk AS
Priority to EP92913614A priority Critical patent/EP0593539A1/fr
Priority to JP5501299A priority patent/JPH06508523A/ja
Publication of WO1993000423A1 publication Critical patent/WO1993000423A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0018—Culture media for cell or tissue culture
    • C12N5/005—Protein-free medium
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/20—Transition metals
    • C12N2500/24—Iron; Fe chelators; Transferrin

Definitions

  • Iron c elate culture medium additive Iron c elate culture medium additive.
  • the present invention relates to an iron supplement for culture media, primarily serum-free or protein-free media, for growing mammalian cells, and a culture medium containing said iron supplement.
  • the present invention relates to a culture medium additive comprising an iron chelate of a soluble iron salt and an alkali metal or alkaline earth metal citrate.
  • Iron chelates for serum-free media have previously been proposed, e.g. in EP 274 445 describing a culture medium additive containing an iron-EDTA/citric acid chelate and aurin tricarboxylic acid.
  • the iron chelate additive of the present invention has the advantage over the one proposed in EP 274 445 that it is composed of inexpensive constituents, and that it contains fewer constituents which might be a ⁇ ource of contamination.
  • the present invention relates to a culture medium for growing mammalian cells, the medium comprising an iron chelate of a soluble iron salt and an alkali metal or alkaline earth metal citrate.
  • the citrate chelator should be mixed with the iron salt so as to generate an equilibrium prior to the addition to the culture medium.
  • This equilibrium may for instance be formed in a concentrated stock solution and, and the process speeded up by stirring, autoclaving, etc.
  • the requisite equilibrium is most conveniently reached when the alkali metal or alkaline earth metal citrate is present in a molar excess relative to the iron salt, in particular a ratio of the citrate to the iron salt of more than 1:1 and less than 500:1.
  • Suitable iron salts for inclusion in the additive of the invention may be selected from the group consisting of FeCl 2 , FeCl 3 , FeS0 4 , Fe 3 (P0 4 ) 2 , Fe(N0 3 ) 3 and Fel 2 .
  • suitable alkali metal or alkaline earth metal citrates for inclusion in the additive of the invention are Na-citrate, K-citrate or Mg- citrate.
  • the iron salt included in the additive is FeCl 2 or FeCl 3
  • the citrate is Na-citrate.
  • a preferred molar ratio of Na-citrate to FeCl 2 /FeCl 3 is between 2:1 and 200:1.
  • the culture medium in which the additive i ⁇ intended to be included is preferably a medium for growing mammalian cells, the additive of the invention constituting an inexpensive iron source which mammalian cells have surprisingly been able to utilise.
  • the medium may for instance be a low-serum medium or, preferably, a serum-free or protein-free medium in which it is important to provide a non-protein iron supplement.
  • mammalian cell ⁇ may al ⁇ o utilise a citrate/iron chloride chelate as the iron source in serum- free media.
  • mammalian cells which exist in an environment enriched in nutrient components and under conditions of considerable osmotic pressure are able to assimilate nutrients in a similar way as a primitive freshwater organism specialized in surviving in a nutrient-poor environment.
  • Adherent BHK cells cultivated in coated T-flask ⁇ containing a serum-free nutrient medium for BHK cells (as described by Maciag et al. 1980, ibid) with transferrin as the only iron source (SFNMT) , were concomitantly inoculated into a series of coated T-flask ⁇ containing serum-free nutrient medium lacking transferrin (SFNM-) but supplemented with a chelated stock solution of Na-citrate and iron chloride.
  • SFNMT transferrin as the only iron source
  • BHK cells were inoculated into spinner flasks containing SFNM- for BHK cells (see example 1) supplemented with a chelated citrate-iron stock solution resulting in 2 mM Citrate and 100 ⁇ M FeCl 3 (final cone.) . Following a few hours where cells were allowed to adhere to coated microcarrier ⁇ , cells spread, propagated and remained es ⁇ entially confluent and healthy for more than two weeks when the experiment was terminated.
  • Adherent CHO cells cultivated in coated T-flasks containing a serum-free nutrient medium for CHO cells (as described by Ham et al. 1979, ibid.) with transferrin as the only iron source (SFNMT) , were concomitantly inoculated into a series of coated T-flask ⁇ containing serum-free nutrient medium lacking transferrin (SFNM-) but supplemented with a chelated stock solution of Na-citrate and iron chloride.
  • SFNMT transferrin as the only iron source
  • Each cell culture was independently treated with respect to replacement of u ⁇ ed medium with fresh serum-free medium of the identical kind or sub-cultivation into new T-flask containing fresh serum-free medium of the identical kind.
  • CHO cells were inoculated into two spinner flasks containing SFNM- for CHO cells (see example 3) supplemented with chelated citrate-iron chloride stock solutions resulting in 2 mM Citrate and 100 and 300 ⁇ M FeCl 3 (final cone), re ⁇ pectively. After a few hour ⁇ where cells were allowed to adhere to coated micro carriers, cells spread, propagated and remained essentially confluent and healthy for more than two weeks when the experiment was terminated.
  • SFNMT transferrin as the only iron source
  • SFNMT transferrin as the only iron source

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

Additif pour milieu de culture comprenant un chélate de fer d'un sel de fer soluble, ainsi qu'un citrate de métal alcalin ou de métal alcalino-terreux. L'additif est une source de fer approprié pour des milieux de culture ne contenant ni sérum ni protéine.
PCT/DK1992/000190 1991-06-21 1992-06-18 Additif pour milieu de culture a base de chelate de fer Ceased WO1993000423A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP92913614A EP0593539A1 (fr) 1991-06-21 1992-06-18 Additif pour milieu de culture a base de chelate de fer
JP5501299A JPH06508523A (ja) 1991-06-21 1992-06-18 鉄キレート培地添加剤

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP91610054.8 1991-06-21
EP91610054 1991-06-21

Publications (1)

Publication Number Publication Date
WO1993000423A1 true WO1993000423A1 (fr) 1993-01-07

Family

ID=8208780

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DK1992/000190 Ceased WO1993000423A1 (fr) 1991-06-21 1992-06-18 Additif pour milieu de culture a base de chelate de fer

Country Status (5)

Country Link
EP (1) EP0593539A1 (fr)
JP (1) JPH06508523A (fr)
AU (1) AU2196892A (fr)
CA (1) CA2111984A1 (fr)
WO (1) WO1993000423A1 (fr)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001016294A3 (fr) * 1999-08-27 2001-09-07 Invitrogen Corp Composes de liaison metallique et leur utilisation dans des compositions de milieu de culture cellulaire
US6767741B1 (en) 1999-08-27 2004-07-27 Invitrogen Corporation Metal binding compounds and their use in cell culture medium compositions
GB2404665A (en) * 2003-08-08 2005-02-09 Cambridge Antibody Tech Culture medium for myeloma cells
WO2005014800A1 (fr) * 2003-08-08 2005-02-17 Cambridge Antibody Technology Limited Culture de cellules de myelome dans un milieu exempt de transferrine et a faible teneur en fer
US7390660B2 (en) 2002-03-05 2008-06-24 Hoffman-La Roche Inc. Methods for growing mammalian cells in vitro
US7598083B2 (en) 2004-10-29 2009-10-06 Centocor, Inc. Chemically defined media compositions
EP2351827A1 (fr) * 2001-03-27 2011-08-03 Life Technologies Corporation Milieu de culture pour la croissance et la transfection de cellules
US9321996B2 (en) 1996-08-30 2016-04-26 Life Technologies Corporation Serum-free mammalian cell culture medium, and uses thereof
EP2250251B1 (fr) 2008-01-09 2017-11-22 Sartorius Stedim Cellca GmbH Additif amélioré de milieu de culture et son procédé d'utilisation
EP3778868A1 (fr) * 2019-08-16 2021-02-17 UGA Biopharma GmbH Milieu de culture cellulaire permettant de cultiver des cellules, procédé de culture des cellules et procédé d'expression d'au moins une protéine recombinant dans une culture cellulaire

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
HK1198657A1 (en) * 2011-10-21 2015-05-22 辉瑞公司 Addition of iron to improve cell culture

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2196348A (en) * 1986-10-03 1988-04-27 Ceskoslovenska Akademie Ved Synthetic medium for hybridoma and myeloma cell cultivation
EP0274445A2 (fr) * 1987-01-09 1988-07-13 Medi-Cult A/S Additif de milieu de culture sans sérum et son utilisation

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2196348A (en) * 1986-10-03 1988-04-27 Ceskoslovenska Akademie Ved Synthetic medium for hybridoma and myeloma cell cultivation
EP0274445A2 (fr) * 1987-01-09 1988-07-13 Medi-Cult A/S Additif de milieu de culture sans sérum et son utilisation

Non-Patent Citations (11)

* Cited by examiner, † Cited by third party
Title
Dialog Information Services, Database BIOSIS, File 5, Dialog Accession No. 9045773, Biosis Accession No. 93030773, FRANEK F.: "Hybridoma growth and monoclonal antibody production in iron-rich protein-free medium effect of nutrient concentration", & Cytotechnology 7 (1), 1991, 33-38. *
Dialog Information Services, File 155, Medline, Dialog Accession No. 02890092, Medline Accession No. 76071092, HILL JH et al.: "Iron-induced enhancement of 67Ga uptake in a model human leukocyte culture system", & J Nucl Med Dec 1975, 16 (12) p1183-6. *
Dialog Information Services, File 155, Medline, Dialog Accession No. 05755034, Medline Accession No. 86056034, REDDEL RR.: "Cell cycle effects of iron depletion on T-47D human breast cancer cells", Exp Cell Res, Dec 1985, 161 (2) p277-84. *
Dialog Information Services, File 155, Medline, Dialog Accession No. 06308060, Medline Accession No. 87282060, HERSHKO C. et al.: "Modification of iron uptake and lipid peroxidation by hypoxia, ascorbic acid, and alpha-tocopherol in iron-loaded rat myocardial cell cultures", & J Lab Clin Med Sep 1987 110 (3) p355-61. *
Dialog Information Services, File 351, WPI, Dialog Accession No. 008681836, WPI Accession No. 91-185855/26, LOEFFLER-INST: "Chemical absorption of ammonia in viral replication medium - by adding iron citrate which reacts to form mixed ligand complex, used in prodn. of antigens for vaccines", DD,A,286612, 910131, 9126 (Basic). *
Dialog Information Services, File 351, WPI, Dialog Accession No. 009027456, WPI Accession No. 92-154816/19, TOSOH CORP: "Complete synthetic medium - contains iron citrate, ethanolamine and linolic acid, oleic acid and/or taurine, does not contain protein, cell growth factor, hormone and steroid"; JP,A,04 091 786, 920325, 9219 *
National Library of Medicine, Database Medline, Accession No. 88284722, KOVAR J.: "Growth-stimulating effect of ferric citrate on hybridoma cells: characterization and relation to transferrin function", & Hybridoma 1988 Jun; 7(3):255-63. *
National Library of Medicine, Database Medline, Accession No. 89124403, SCHNEIDER Y.: "Optimisation of hybridoma cell growth and monoclonal antibody secretion in a chemically defined, serum- and protein-free culture medium", & J Immunol Methods 1989 Jan 6;116(1):65-77. *
PATENT ABSTRACTS OF JAPAN, Vol. 12, No. 209, C504; & JP,A,63 077 801, publ 1988-01-13 Nippon Zenyaku Kogyo K.K. *
PATENT ABSTRACTS OF JAPAN, Vol. 12, No. 398, C538; & JP,A,63 141 584, publ 1988-06-14 Chemo Sero Therapeut Res Inst. *
Tibtech, Vol. 7, September 1989 E. SHACTER: "Serum-free media for bulk culture of hybridoma cells and the preparation of monoclonal antibodies", see page 248 - page 253, see page 249, right column. *

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9321996B2 (en) 1996-08-30 2016-04-26 Life Technologies Corporation Serum-free mammalian cell culture medium, and uses thereof
US6767741B1 (en) 1999-08-27 2004-07-27 Invitrogen Corporation Metal binding compounds and their use in cell culture medium compositions
WO2001016294A3 (fr) * 1999-08-27 2001-09-07 Invitrogen Corp Composes de liaison metallique et leur utilisation dans des compositions de milieu de culture cellulaire
US9879243B2 (en) 2001-03-27 2018-01-30 Lifetechnologies Corporation Culture medium for cell growth and transfection
EP2351827A1 (fr) * 2001-03-27 2011-08-03 Life Technologies Corporation Milieu de culture pour la croissance et la transfection de cellules
US7390660B2 (en) 2002-03-05 2008-06-24 Hoffman-La Roche Inc. Methods for growing mammalian cells in vitro
GB2404665A (en) * 2003-08-08 2005-02-09 Cambridge Antibody Tech Culture medium for myeloma cells
JP2007501606A (ja) * 2003-08-08 2007-02-01 ケンブリッジ アンチボディー テクノロジー リミテッド トランスフェリン不含・低鉄培地中での骨髄腫細胞培養
GB2404665B (en) * 2003-08-08 2005-07-06 Cambridge Antibody Tech Cell culture
US8361797B2 (en) 2003-08-08 2013-01-29 Medimmune Limited Myeloma cell culture in transferrin-free low iron medium
WO2005014800A1 (fr) * 2003-08-08 2005-02-17 Cambridge Antibody Technology Limited Culture de cellules de myelome dans un milieu exempt de transferrine et a faible teneur en fer
US7598083B2 (en) 2004-10-29 2009-10-06 Centocor, Inc. Chemically defined media compositions
EP2250251B1 (fr) 2008-01-09 2017-11-22 Sartorius Stedim Cellca GmbH Additif amélioré de milieu de culture et son procédé d'utilisation
EP3778868A1 (fr) * 2019-08-16 2021-02-17 UGA Biopharma GmbH Milieu de culture cellulaire permettant de cultiver des cellules, procédé de culture des cellules et procédé d'expression d'au moins une protéine recombinant dans une culture cellulaire
WO2021032637A1 (fr) * 2019-08-16 2021-02-25 Uga Biopharma Gmbh Milieu de culture cellulaire pour la culture de cellules, procédé de culture cellulaire et procédé d'expression d'au moins une protéine recombinée dans une culture cellulaire
US20220298472A1 (en) * 2019-08-16 2022-09-22 Uga Biopharma Gmbh Cell culture medium for cultivating cells, method for cultivating cells, and method for expressing at least one recombinant protein in a cell culture
JP2022544800A (ja) * 2019-08-16 2022-10-21 ウーゲーアー バイオファーマ ゲーエムベーハー 細胞を培養するための細胞培養培地、細胞培養方法および細胞培養における少なくとも１つの組換えタンパク質の発現方法
US12421492B2 (en) 2019-08-16 2025-09-23 Uga Biopharma Gmbh Cell culture medium for cultivating cells, method for cultivating cells, and method for expressing at least one recombinant protein in a cell culture

Also Published As

Publication number Publication date
CA2111984A1 (fr) 1993-01-07
JPH06508523A (ja) 1994-09-29
AU2196892A (en) 1993-01-25
EP0593539A1 (fr) 1994-04-27

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