EP0593539A1 - Additif pour milieu de culture a base de chelate de fer - Google Patents
Additif pour milieu de culture a base de chelate de ferInfo
- Publication number
- EP0593539A1 EP0593539A1 EP92913614A EP92913614A EP0593539A1 EP 0593539 A1 EP0593539 A1 EP 0593539A1 EP 92913614 A EP92913614 A EP 92913614A EP 92913614 A EP92913614 A EP 92913614A EP 0593539 A1 EP0593539 A1 EP 0593539A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- citrate
- culture medium
- fecl
- iron
- iron salt
- 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- Withdrawn
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
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.
Priority Applications (1)
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 |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP91610054 | 1991-06-21 | ||
EP91610054 | 1991-06-21 | ||
PCT/DK1992/000190 WO1993000423A1 (fr) | 1991-06-21 | 1992-06-18 | Additif pour milieu de culture a base de chelate de fer |
EP92913614A EP0593539A1 (fr) | 1991-06-21 | 1992-06-18 | Additif pour milieu de culture a base de chelate de fer |
Publications (1)
Publication Number | Publication Date |
---|---|
EP0593539A1 true EP0593539A1 (fr) | 1994-04-27 |
Family
ID=8208780
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP92913614A Withdrawn EP0593539A1 (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) |
Families Citing this family (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2243827B2 (fr) | 1996-08-30 | 2017-11-22 | Life Technologies Corporation | Milieu de culture de cellules de mammiferes exempt de serum |
CA2383460C (fr) * | 1999-08-27 | 2015-10-13 | Invitrogen Corporation | 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 |
US20030096414A1 (en) * | 2001-03-27 | 2003-05-22 | Invitrogen Corporation | Culture medium for cell growth and transfection |
CA2417689C (fr) | 2002-03-05 | 2006-05-09 | F. Hoffmann-La Roche Ag | Methodes ameliorees de culture in vitro de cellules de mammiferes |
DE602004005868T2 (de) * | 2003-08-08 | 2007-12-20 | Cambridge Antibody Technology Ltd. | Myeloma zellkultur in einem transferrin-freien medium mit niedrigem eisengehalt |
GB2404665B (en) | 2003-08-08 | 2005-07-06 | Cambridge Antibody Tech | Cell culture |
WO2006050050A2 (fr) | 2004-10-29 | 2006-05-11 | Centocor, Inc. | Compositions de milieux chimiquement definis |
DK2250251T3 (en) | 2008-01-09 | 2018-01-22 | Sartorius Stedim Cellca Gmbh | Improved culture medium additive and method of using it |
AU2012324495B2 (en) * | 2011-10-21 | 2016-04-21 | Pfizer Inc. | Addition of iron to improve cell culture |
EP3778868B1 (fr) * | 2019-08-16 | 2022-01-26 | 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 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CS262822B1 (en) * | 1986-10-03 | 1989-04-14 | Kovar Jan | Synthetic medium for the cultivation of myelomic cells |
NO162160C (no) * | 1987-01-09 | 1989-11-15 | Medi Cult As | Serumfritt vekstmedium, samt anvendelse derav. |
-
1992
- 1992-06-18 JP JP5501299A patent/JPH06508523A/ja active Pending
- 1992-06-18 CA CA 2111984 patent/CA2111984A1/fr not_active Abandoned
- 1992-06-18 WO PCT/DK1992/000190 patent/WO1993000423A1/fr not_active Application Discontinuation
- 1992-06-18 EP EP92913614A patent/EP0593539A1/fr not_active Withdrawn
- 1992-06-18 AU AU21968/92A patent/AU2196892A/en not_active Abandoned
Non-Patent Citations (1)
Title |
---|
See references of WO9300423A1 * |
Also Published As
Publication number | Publication date |
---|---|
WO1993000423A1 (fr) | 1993-01-07 |
JPH06508523A (ja) | 1994-09-29 |
AU2196892A (en) | 1993-01-25 |
CA2111984A1 (fr) | 1993-01-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP0274445B1 (fr) | Additif de milieu de culture sans sérum et son utilisation | |
US5122469A (en) | Method for culturing Chinese hamster ovary cells to improve production of recombinant proteins | |
Jan et al. | Peptone, a low-cost growth-promoting nutrient for intensive animal cell culture | |
Iype | Cultures from adult rat liver cells I. Establishment of monolayer cell‐cultures from normal liver | |
EA023193B1 (ru) | Среда для культивирования клеток для экспрессии белков adamts | |
CN101418330B (zh) | 适于nso细胞大规模培养及抗体生产的无蛋白培养基 | |
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 | |
JP2015062436A (ja) | 連続細胞培養で対象となるポリペプチドまたはウイルスを生成する方法 | |
CN1962857A (zh) | 一种哺乳动物细胞无血清培养基 | |
JP2016052310A (ja) | 改良型細胞培養培地 | |
JPH06511389A (ja) | 動物細胞培養 | |
EP0593539A1 (fr) | Additif pour milieu de culture a base de chelate de fer | |
CN102317440A (zh) | 改进的培养基添加剂及其应用方法 | |
CN108925137A (zh) | 细胞培养基 | |
CN106399224A (zh) | 无血清无蛋白细胞培养基 | |
Minamoto et al. | Development of a serum-free and heat-sterilizable medium and continuous high-density cell culture | |
JPH08508875A (ja) | 蛋白分泌細胞の流加回分培養法 | |
JPH09512171A (ja) | 無血清培地添加物 | |
NZ229187A (en) | Method of production of t-pa from cell culture | |
JP2016508376A (ja) | 組換えタンパク質産生の増加のための製剤及び方法 | |
JPH03254679A (ja) | 新規細胞株 | |
JPH0432078B2 (fr) | ||
WO1988001644A1 (fr) | Procede d'isolement de produits secretes par une tumeur en utilisant un nouveau milieu exempt de proteines | |
US5602025A (en) | Non-tumorigenic cell lines for expression of genes | |
JP2001029066A (ja) | ヒト・動物細胞培養液 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 19931208 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AT BE CH DE DK ES FR GB GR IT LI LU NL SE |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
18D | Application deemed to be withdrawn |
Effective date: 19980102 |