[go: up one dir, main page]

WO1993000423A1 - Iron chelate culture medium additive - Google Patents

Iron chelate culture medium additive Download PDF

Info

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
French (fr)
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 JP5501299A priority Critical patent/JPH06508523A/en
Priority to EP92913614A priority patent/EP0593539A1/en
Publication of WO1993000423A1 publication Critical patent/WO1993000423A1/en
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

Landscapes

  • 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

A culture medium additive comprises an iron chelate of a soluble iron salt and an alkali metal or alkaline earth metal citrate. The additive is a suitable iron source for serum-free or protein-free culture media.

Description

Iron c elate culture medium additive.
FIELD OF INVENTION
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.
BACKGROUND OF THE INVENTION
Until fairly recently, conventional media for growing mammalian cells contained serum as an important source of growth factors in the requisite concentrations for the growth and natural multiplication of the cells. The presence of serum or specific added proteins in culture media, however, suffers from the disadvantage that the purification of the desired protein product from the mammalian culture is made more difficult and that there is an increased risk of contamination by infectious agents. It is therefore an important aim in the field of mammalian cell culture to develop media in which the components in serum necessary for cell growth have been replaced with non- proteinaceous substances serving the same purpose. Serum-free or protein-free media have therefore become increasingly important for the cultivation of mammalian cells in the production of biological materials (e.g. monoclonal antibodies, natural or recombinant pharmaceuticals, or the like) .
Most serum-free media are based on a commercially available basal medium (e.g. MEM, Ham, RPMI) supplemented with insulin, transferrin, selenium, growth factors, and some protein and lipid sources \Hamilton et al. , In Vitro 13: 537-547, 1977; Ham et al. , Methods Enzymol. 58: 44-93, 1979; Maciag et al.. Cell Biol. Int. Rep. 4: 43-50, 1980; Barnes, BioTechnology 5: 534- 540, 1987; Fiorentini et al. , Am. Biotech. Lab. 8: 35-37, 1990; Bjare, J. Biotech. 1.5: 147-154, 1990; Hewlett, Cytotechnology 5: 3-14, 1991]. SUMMARY OF THE INVENTION
It has now been found possible to replace transferrin as the iron source in serum-free media by a non-protein chelate of citrate and an iron salt.
Accordingly, 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 εource of contamination.
In another aspect, 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.
DETAILED DISCLOSURE OF THE INVENTION
To avoid iron precipitation and potential toxic effects of the iron on the cultured cells, 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. In the preparation of the iron additive, 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 FeCl2, FeCl3, FeS04, Fe3(P04)2, Fe(N03)3 and Fel2. Examples of suitable alkali metal or alkaline earth metal citrates for inclusion in the additive of the invention are Na-citrate, K-citrate or Mg- citrate. In a particularly preferred embodiment, the iron salt included in the additive is FeCl2 or FeCl3, and the citrate is Na-citrate. In this case, a preferred molar ratio of Na-citrate to FeCl2/FeCl3 is between 2:1 and 200:1.
The culture medium in which the additive iε 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. Thus, 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. Although it has previouεly been deεcribed that the freshwater ciliate Tetrahy ena thermophila is able to utilise pre-chelated iron citrate as the only iron source (cf. P.B. Suhr-Jessen and L. Raεmusεen, Exp. Cell Res. 139. 1982, pp. 457-460; L. Ras ussen et al., J. Cell. Phys. 122. 1985, pp. 155-158), it haε not been suggeεted that mammalian cellε may alεo utilise a citrate/iron chloride chelate as the iron source in serum- free media. Biologically speaking, it is quite surprising that 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.
The invention is further illustrated in the following examples which are not in any way intended to limit the scope of the invention as claimed. EXAMPLE 1. BHK cells
Adherent BHK cells cultivated in coated T-flaskε 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-flaskε containing serum-free nutrient medium lacking transferrin (SFNM-) but supplemented with a chelated stock solution of Na-citrate and iron chloride. Experiments 1 to 3 had different durations and the experimental citrate concentration was 2 mM, 2 mM, and 5 mM (final cone), respectively. Parallel control cultures were cultivated in SFNMT.
Each cell culture was independently treated with respect to replacement of used medium with freεh serum-free medium of the identical kind or εub-cultivation into new T-flask containing fresh serum-free medium of the identical kind. At the end of the experiment, the total number of doublings in each medium was calculated:
Figure imgf000006_0001
Citrate and iron chloride was not added to SFNMT EXAMPLE 2. BHK cells
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 μM FeCl3 (final cone.) . Following a few hours where cells were allowed to adhere to coated microcarrierε, cells spread, propagated and remained esεentially confluent and healthy for more than two weeks when the experiment was terminated.
EXAMPLE 3. CHO cells
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-flaskε containing serum-free nutrient medium lacking transferrin (SFNM-) but supplemented with a chelated stock solution of Na-citrate and iron chloride. Experiments 1 and 2 had different durations and the experimental citrate concentration was 2 mM (final cone.) . Parallel control cultures were cultivated in SFNMT.
Each cell culture was independently treated with respect to replacement of uεed 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.
At the end of the experiment, the total number of doublings in each medium was calculated:
Figure imgf000008_0001
Citrate and iron chloride was not added to SFNMT
EXAMPLE 4. CHO cells
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 μM FeCl3 (final cone), reεpectively. After a few hourε 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.
EXAMPLE 5. MYELOMA cells
SP2/0 myeloma cells cultivated in suspension culture in T- flaskε containing an RPMI baεed serum-free nutrient medium (Shacter 1989, TIBTECH, 7, 248-253) with transferrin as the only iron source (SFNMT) , were concomittantly inoculated into a series of T-flasks containing serum-free nutrient medium lacking transferrin (SFNM-) but supplemented with a chelated stock εolution of Na-citrate and iron chloride.
Each cell culture was independently treated with respect to replacement of used medium with fresh serum-free medium of the identical kind or sub-cultivation into new T-flask containing freεh serum-free medium of the identical kind. At the end of the experiment, the total number of doublings in each medium waε calculated:
Figure imgf000009_0001
Citrate and iron chloride was not added to SFMNT
EXAMPLE 6. HYBRIDOMA cells
SP2/0 based hybridoma cells cultivated in suεpenεion culture in T-flaεks containing an RPMI based serum-free nutrient medium for hybridoma cells (Shacter 1989, TIBTECH, 7, 248-253) with transferrin as the only iron source (SFNMT) , were concomittantly inoculated into a series of T-flasks containing serum-free nutrient medium lacking transferrin (SFNM-) but supplemented with a chelated stock εolution of Na-citrate and iron chloride.
Each cell culture waε independently treated with reεpect to replacement of uεed 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. At the end of the experiment, the total number of doublingε in each medium was calculated:
Figure imgf000010_0001
Citrate and iron chloride was not added to SFNMT

Claims

1. An culture medium additive comprising an iron chelate of a soluble iron salt and an alkali metal or alkaline earth metal citrate.
2. An additive according to claim 1, wherein the alkali metal or alkaline earth metal citrate iε present in a molar excess relative to the iron salt
3. An additive according to claim 1 or 2, wherein the iron salt is εelected from the group conεisting of FeCl2, FeCl3, FeS04, Fe3(P04)2, Fe(N03)3 and Fel2.
4. An additive according to any of claims 1-3, wherein the alkali metal or alkaline earth metal citrate iε selected from the group consisting of Na-citrate, K-citrate and Mg-citrate.
5. An additive according to any of claims 1-4, wherein the molar ratio of alkali metal or alkaline earth metal citrate to iron salt is more than 1:1 and less than 500:1.
6. An additive according to any of claimε 1-5, wherein the culture medium in which it iε included is for growing mammalian cells.
7. An additive according to any of claims 1-6, wherein the culture medium in which it is included iε a serum-free or protein-free medium.
8. An additive according to any of claims 1-7, wherein the iron salt is FeCl2 or FeCl3, and wherein the citrate is Na-citrate.
9. An additive according to claim 8, wherein the molar ratio of Na-citrate to FeCl2/FeCl3 is between 2:1 and 200:1.
10. 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.
11. A culture medium according to claim 10, wherein the alkali metal or alkaline earth metal citrate is present in a molar excess relative to the iron salt.
12. A culture medium according to claim 10 or 11, wherein the iron salt is selected from the group consisting of FeCl2, FeCl3, FeS04, Fe3(P04)2, Fe(N03)3 and Fel2.
13. A culture medium according to any of claims 10-12, wherein the alkali metal or alkaline earth metal citrate is selected from the group consisting of Na-citrate, K-citrate and Mg- citrate.
14. A culture medium according to any of claimε 10-13, wherein the molar ratio of alkali metal or alkaline earth metal citrate to iron salt is more than 1:1 and less than 500:1.
15. A culture medium according to any of claims 10-14, which is a serum-free or protein-free medium.
16. A culture medium according to any of claims 10-15, wherein the iron salt is FeCl2 or FeCl3, and wherein the citrate is Na- citrate.
17. A culture medium according to claim 16, wherein the molar ratio of Na-citrate to FeCl2/FeCl3 is between 2:1 and 200:1.
PCT/DK1992/000190 1991-06-21 1992-06-18 Iron chelate culture medium additive Ceased WO1993000423A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP5501299A JPH06508523A (en) 1991-06-21 1992-06-18 Iron chelate medium additive
EP92913614A EP0593539A1 (en) 1991-06-21 1992-06-18 Iron chelate culture medium additive

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 (en) 1993-01-07

Family

ID=8208780

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DK1992/000190 Ceased WO1993000423A1 (en) 1991-06-21 1992-06-18 Iron chelate culture medium additive

Country Status (5)

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

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001016294A3 (en) * 1999-08-27 2001-09-07 Invitrogen Corp Metal binding compounds and their use in cell culture medium compositions
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 (en) * 2003-08-08 2005-02-17 Cambridge Antibody Technology Limited Myeloma cell culture in transferrin-free low iron medium
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 (en) * 2001-03-27 2011-08-03 Life Technologies Corporation Culture medium for cell growth and transfection
US9321996B2 (en) 1996-08-30 2016-04-26 Life Technologies Corporation Serum-free mammalian cell culture medium, and uses thereof
EP2250251B1 (en) 2008-01-09 2017-11-22 Sartorius Stedim Cellca GmbH Improved culture media additive and process for using it
EP3778868A1 (en) * 2019-08-16 2021-02-17 UGA Biopharma GmbH Cell culture medium for cultivating cells, method for culturing cells and method for expressing at least one recombinant protein in a cell culture

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2852021C (en) * 2011-10-21 2018-08-28 Pfizer Inc. 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 (en) * 1987-01-09 1988-07-13 Medi-Cult A/S A serum-free growth medium additive and a use thereof

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 (en) * 1987-01-09 1988-07-13 Medi-Cult A/S A serum-free growth medium additive and a use thereof

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 (en) * 1999-08-27 2001-09-07 Invitrogen Corp Metal binding compounds and their use in cell culture medium compositions
US9879243B2 (en) 2001-03-27 2018-01-30 Lifetechnologies Corporation Culture medium for cell growth and transfection
EP2351827A1 (en) * 2001-03-27 2011-08-03 Life Technologies Corporation Culture medium for cell growth and transfection
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 (en) * 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 Myeloma cell culture in transferrin-free and low iron medium
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 (en) * 2003-08-08 2005-02-17 Cambridge Antibody Technology Limited Myeloma cell culture in transferrin-free low iron medium
US7598083B2 (en) 2004-10-29 2009-10-06 Centocor, Inc. Chemically defined media compositions
EP2250251B1 (en) 2008-01-09 2017-11-22 Sartorius Stedim Cellca GmbH Improved culture media additive and process for using it
EP3778868A1 (en) * 2019-08-16 2021-02-17 UGA Biopharma GmbH Cell culture medium for cultivating cells, method for culturing cells and method for expressing at least one recombinant protein in a cell culture
WO2021032637A1 (en) * 2019-08-16 2021-02-25 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
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 (en) * 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 Cell culture media for culturing cells, cell culture methods and methods for expressing at least one recombinant protein in cell culture
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
EP0593539A1 (en) 1994-04-27
AU2196892A (en) 1993-01-25
CA2111984A1 (en) 1993-01-07
JPH06508523A (en) 1994-09-29

Similar Documents

Publication Publication Date Title
US5045454A (en) Serum-free growth medium and use thereof
Jan et al. Peptone, a low-cost growth-promoting nutrient for intensive animal cell culture
Prasad et al. Vitamin K3 (menadione) inhibits the growth of mammalian tumor cells in culture
US20150329830A1 (en) Myeloma cell culture in transferrin-free low iron medium
Heath et al. Methods for increasing monoclonal antibody production in suspension and entrapped cell cultures: biochemical and flow cytometric analysis as a function of medium serum content
EA023193B1 (en) Cell culture medium for adamts protein expression
CN101418330B (en) Non protein culture medium adapted to large-scale culture of NSO cell and production of antibody
WO2008009642A1 (en) Cell culture media
CN102317440A (en) Improved culture medium additive and application method thereof
WO2006047380A2 (en) Method and media for single cell serum-fee culture of cho cells
WO1993000423A1 (en) Iron chelate culture medium additive
EP1210410B1 (en) Metal binding compounds and their use in cell culture medium compositions
Minamoto et al. Development of a serum-free and heat-sterilizable medium and continuous high-density cell culture
Shintani et al. Polyethylene glycols for promoting the growth of mammalian cells
CN106399224A (en) Serum-free and protein-free cell culture medium
US5871999A (en) Cell culture process and medium for the growth of adherent animal cells
EP0248656A2 (en) Composition for cell cultivation and use thereof
JPH09512171A (en) Serum-free medium additive
JP2859679B2 (en) New cell line
US4879222A (en) Method of isolating tumor-secreted products using a novel protein-free medium
JP2016508376A (en) Formulations and methods for increasing recombinant protein production
Keenan et al. Replacement of transferrin by simple iron compounds for MDCK cells grown and subcultured in serum-free medium
US5602025A (en) Non-tumorigenic cell lines for expression of genes
US6858428B2 (en) Human and animal cell culture medium containing antithrombin III for suppressing cell death
JP2850430B2 (en) Serum-free medium for cell culture

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IT LU MC NL SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 1992913614

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2111984

Country of ref document: CA

ENP Entry into the national phase

Ref document number: 1994 167891

Country of ref document: US

Date of ref document: 19940105

Kind code of ref document: A

WWP Wipo information: published in national office

Ref document number: 1992913614

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1992913614

Country of ref document: EP