Abstract
A serum free medium was developed, that could be used for the large scale propagation of various cell lines in bioreactors. The medium is based on a 1:1 mixture of Iscove's Modified Dulbecco's Medium and Ham's Medium F12, supplemented with transferrin, insulin and a BSA/oleic acid complex. Several myelomas, hybridomas derived from different myelomas and spleen cells, and other lymphoid and non-lymphoid cell lines were cultivated at growth rates comparable to those observed using serum-supplemented media. There was furthermore no reduction in the formation of products such as monoclonal antibodies or recombinant human interleukin-2.
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Abbreviations
- Ag8:
-
Mouse myeloma cell line P3-X63-Ag8.653
- BME:
-
Basal Medium Eagle
- BSA:
-
Bovine Serum Albumin
- DMEM:
-
Dulbecco's Modified Eagle's Medium
- EDTA:
-
Ethylenediaminete-traacetic Acid
- e-PC:
-
Phosphatidyl choline from egg yolk
- FCS:
-
Fetal Calf Serum
- FGF:
-
Fibroblast Growth Factor
- GHL:
-
Glycyl-histidyl-lysine
- HDL:
-
High Density Lipoprotein
- HPL:
-
Human Plasma Lipid
- IF:
-
1:1 mixture of IMDM and Ham's F12
- IMDM:
-
Iscove's Modified Dulbecco's medium
- LDL:
-
Low Density Lipoprotein
- NS1:
-
Mouse myeloma cell line NSI-1-Ag4-1
- PBS:
-
Phosphate Buffered Saline
- s-PC:
-
Phosphatidylcholine from soy beans
- s-PE:
-
Phosphatidylethanolamine from soy beans
- s-lecithin:
-
lecithin from soy beans
References
Büntemeyer H, Bödeker BGD and Lehmann J (1987) Membrane-stirrer-reactor for bubble-free aeration and perfusion. In: Spier RE, Griffiths JB (eds) Modern Approaches to Animal Cell Technology (pp. 411–419) Butterworth & Co. Ltd.
Büntemeyer H (1987) Entwicklung eines Perfusionssystems zur Kultivierung tierischer Zellen in Suspension. Thesis, University of Hannover, FRG.
Chang TH, Steplewski Z and Koprowski H (1980) Production of monoclonal antibodies in serum free medium. J. Immunol. Methods 39: 369–375.
Cleveland WL, Wood I and Erlanger BF (1983) Routine large-scale production of monoclonal antibodies in a protein-free culture medium. J. Immunol. Methods 56: 221–234.
Conradt HS, Ausmeier M, Dittmar KEJ, Hauser H and Lindenmaier W (1986) Secretion of glycosylated human II-2 by recombinant mammalian cell lines. Carbohydrate Res. 149: 443–450.
Grillis S, Ferm MM, Ou W and Smith KA (1978) T cell growth factor: Parameters of production and quantitative microassay for activity. J. Immunol. 120: 2027–2032.
Jäger V, Büntemeyer H and Lehmann J (1987) Increasing the rate of production of monoclonal antibodies by combining a stirred reactor with a hollow fiber perfusion system. In: Neijssel OM, van der Meer RR, Luyben KChAM (eds) Proc. 4th Europ. Congr. on Biotechnol. Vol. 3 (pp. 581–584) Elsevier Pub.
Kawamoto T, Sato JD, Le A, McClure DB and Sato GH (1983) Development of a serum-free medium for growth of NS-1 mouse myeloma cells and its application to the isolation of NS-1 hybridomas. Anal. Biochem. 130: 445–453.
Kovar J and Franek F (1984) Serum free medium for hybridoma and parental myeloma cell growth cultivation: A novel composition of growth-supporting substances. Immunology Letters 7: 339–345.
Lehmann J, Piehl GW, Schulz R (1987) Bubble-free cell culture aeration with porous moving membranes. Dev Biol. Standard. 66: 227–240.
Murakami H, Masui H, Sato GH and Raschke WC (1981) Growth of mouse plasmacytoma cells in serum-free, hormone-supplemented medium: Procedure of the determination of hormone and growth factor requirements for cell growth. Anal. Biochem. 114: 422–428.
Murakami H, Edamoto T, Nakamura H and Omura H (1982) Growth of MPC-11 cells in serum-free growth factor supplemented medium. Agric. Biol. Chem. 46: 1831–1837.
Murakami H, Masui H and Sato GH (1982) Suspension cultures of hybridoma cells in serum-free medium: Soybean phospholipids as the essential components. In: Sato GH, Pardee AB, Sirbasku DA (eds) Growth of Cells in Hormonally Defined Media (pp. 711–715) Cold Springs Harbor Laboratory Press.
Murakami H, Masui H, Sato GH, Sueoka N, Chow TP and Kano-Sueoka T (1982) Growth of hybridoma cells in serum-free medium: Ethanolamine is an essential component. Proc. Natl. Acad. Sci. USA 79: 1158–1162.
Pickart L, Thayer L and Thaler M (1973) A synthetic tripeptide which increases survival of normal liver cells, and stimulates growth in hepatoma cells. Biochem. Biophys. Res. Commun. 54: 562–566.
Reuveny S, Velez D, Miller L and Macmillan JD (1986) Comparison of cell propagation methods for their effect on monoclonal antibody yield in fermentors. J. Immunol. Methods 86: 61–69.
Samoilovich SR, Dugan CB and Macario AJL (1987) Hybridoma technology: New developments of practical interest. J. Immunol. Methods 101: 153–170.
Tauber J-P, Cheng J, Massoglia S and Gospodarowicz D (1981) High density lipoproteins and the growth of vascular endothelial cells in serum-free medium. In Vitro 17: 519–530.
Yamane I (1978) Role of bovine albumin in a serum-free culture medium and its application. Natl. Cancer Inst. Monogr. 48: 131–133.
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Jäger, V., Lehmann, J. & Friedl, P. Serum-free growth medium for the cultivation of a wide spectrum of mammalian cells in stirred bioreactors. Cytotechnology 1, 319–329 (1988). https://doi.org/10.1007/BF00365077
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DOI: https://doi.org/10.1007/BF00365077