Abstract
Suppression of proliferation of cells which contain stable or stabilized mRNA coded for a protein to be produced, a partial mimic of cell differentiation, was examined for enhancing protein production by cultured mammalian cells. Hybridoma 2E3 cells which were adapted to be interleukin-6 sensitively growth-suppressed accumulated the mRNA of IgG1 which is reported stable, and IgG1 production rate increased as a result when their growth was suppressed with interleukin-6. A myeloma cell line was similarly adapted; the obtained myeloma cells can be used as host cells for enhancing production of exogenous proteins by suppressing growth with interleukin-6. Temperature-sensitively growth-suppressible mutants of mouse mammary carcinoma FM3A were transfected with cDNA of IgM λ1 chain and cultured at nonpermissive temperature to enhance production of λ1. Addition of various growth-suppressive reagents to culture medium was studied for finding methods suitable for suppressing growth while maintaining high cell viability. Caffeine yielded the best results among these reagents. Deprivation of various growth-supporting components in culture medium was also tested; simultaneous deprivation of insulin and transferrin viably suppressed growth of hybridoma 2E3 cells, resulting in enhanced antibody productivity.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Abbreviations
- IL6:
-
recombinant human interleukin-6
- TGF:
-
β-recombinant human TGF-β1
- X63.653-P3X63:
-
Ag8.653 myeloma
References
Brock ML and Shapiro DJ (1983) Estrogen stabilizes vitellogenin mRNA against cytoplasmic degradation. Cell 34: 207–214.
Caput D, Beutler B, Hartog K, Thayer R, Brown-Shimer S and Cerami A (1986) Identification of a common nucleotide sequence in the 3′ untranslated region of mRNA molecules specifying inflammatory mediators. Proc. Natl. Acad. Sci. USA 83: 1670–1674.
Carneiro M and Schibier U (1984) Accumulation of rare and moderately abundant mRNAs in mouse L-cells is mainly post-transcriptionally regulated. J. Mol. Biol. 178: 869–880.
Darzynkiewicz Z, Traganos F, Xue SH and Melamed MR (1981) Effect of n-butyrate on cell cycle progression and in situ chromatin structure of L1210 cells. Exp. Cell Res. 136: 279–293.
Kafatos FC (1972) The cocoonase zymogen cells of silk moths: a model of terminal cell differentiation for specific protein synthesis. Current Topics in Developmental Biology 7: 125–191.
Karasuyama H, Kudo A and Melchers F (1990) The proteins encoded by the VpreB and λ5 pre-B cell-specific genes can associate with each other and with μ heavy chain. J. Exp. Med. 172: 969–972.
Kashima N, Nishi-Takaoka C, Fujita T, Taki S, Yamada G, Hamuro J and Taniguchi T (1985) Unique structure of murine interleukin-2 as deduced from cloned cDNAs. Nature 313: 402–404.
Koeller, DM, Horowitz JA, Casey JL, Klausner RD and Harford JB (1991) Translational and the stability of mRNAs encoding the transferrin receptor and c-fos. Proc. Natl. Acad. Sci. USA 88: 7778–7782.
Kuehl WM (1977) Synthesis of immunoglobulin in myeloma cells. Curr. Top. Microbiol. Immunol. 76: 1–46.
Makishima F, Terada S, Mikami T and Suzuki E (1992) Interleukin-6 is antiproliferative to a mouse hybridoma cell line and promotive for its antibody productivity. Cytotechnol. 10: 15–23.
Murakami F, Yasuda H, Miyazawa H, Hanaoka F and Yamada M (1985) Characterization of a temperature-sensitive mutant of mouse FM3A cells defective in DNA replication. Proc. Natl. Acad. Sci. USA 82: 1761–1765.
Okuda A and Kimura G (1988) Elongation of Gl phase by transient exposure of rat 3Y1 fibroblasts to caffeine during the previous and present generations. J. Cell Sci. 89: 379–386.
Savinell JM, Lee GM and Palsson BO (1989) On the orders of magnitude of epigenic dynamics and monoclonal antibody production. Bioprocess Eng. 4: 231–234.
Suzuki E (1990) Kinetics of monoclonbal antibody production. Doctoral dissertation, North Carolina State University, Raleigh, North Carolina.
Suzuki E and Ollis DF (1990) Enhanced antibody production at slowed growth rates: experimental demonstration and a simple structured model. Biotechnol. Prog. 6: 231–236.
Suzuki E, Takahashi K and Ollis DF (1992) A simple structured model predicted positively-, negatively, or non-growth associated antibody production rate depending on culture conditions. In: Murakami H, Shirahata S and Tachibana H (eds.) Animal Cell Technology: Basic and Applied Aspects vol. 4 (pp. 375–381).
Storb U (1973) Turnover of myeloma messenger RNA. Biochem. Biophys. Res. Commun. 52: 1483–1491.
Traganos F, Darzynkiewicz Z and Melamed MR (1982) The ratio of RNA to total nucleic acid content as a quantitative measure of unbalanced cell growth. Cytometry 2: 212–218.
Yasuda H, Kamijo M, Honda R, Nakamura M, Hanaoka F and Ohba Y (1991) A point mutation in C-terminal region of cdc2 kinase causes a G2-phase arrest in a mouse temperature-sensitive FM3 A cell mutant. Cell Struct. Funct. 16: 105–112.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1994 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Takahashi, K., Tereda, S., Ueda, H., Makishima, F., Suzuki, E. (1994). Growth rate suppression of cultured mammalian cells enhances protein productivity. In: Buckland, B.C., Aunins, J.G., Bibila, T.A., Hu, WS., Robinson, D.K., Zhou, W. (eds) Cell Culture Engineering IV. Current Applications of Cell Culture Engineering, vol 1. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-0257-5_7
Download citation
DOI: https://doi.org/10.1007/978-94-011-0257-5_7
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-4114-0
Online ISBN: 978-94-011-0257-5
eBook Packages: Springer Book Archive