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Establishment of Namalva cell lines which grow continuously in glutamine-free medium

Cytotechnology volume 1, pages 151–158 (1988)Cite this article

Abstract

Glutamine has been shown to be a preferred energy source for some established cell lines and cancer cells in culture (Kovacevic, 1971; Kovacevic, 1972; Lavietes, 1974). Empirically, glutamine is the most abundant amino acid in most of the culture media developed. The major end product of glutamine metabolism is ammonia. Ammonia build up is one of the limiting factors in the proliferation of mammalian cells in higher density culture and is directly related to the initial glutamine concentration. The susceptibility of glutamine to thermodecomposition prevents the heat sterilization of glutamine-enriched media and this significantly increases the cost of medium preparation at the industrial scale. In an attempt to overcome these drawbacks, a population of Namalva cells capable of growing in glutamine-free media was established. The adapted cells were found to contain a higher level of glutamine synthetase activity which enable them to synthesize sufficient amounts of glutamine for their growth.

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Abbreviations

GS:

glutamine synthetase

HEPES:

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

References

  1. Butler M (1985) Growth limitations in high density micro-carrier cultures. Develop. Biol. Standard. 60: 269–280.

    Google Scholar 

  2. Butler M, Imamura T, Thomas J and Thilly WG (1983) High yields from microcarrier cultures by medium perfusion. J. Cell Sci. 61: 351–363.

    Google Scholar 

  3. Eagle H, Washington CL, Ievy M and Cohen L (1966) The Population-dependent Requirement by Cultured Mammalian Cells for Metabolites Which They Can Synthesize. II. Glutamic acid and Glutamine; Aspartic acid and Asparagine. J. Biol. Chem. 241: 4994–4999.

    Google Scholar 

  4. Hayward BE, Hussain A, Wilson RH, Lyons A, Woodcock V, McIntosh B and Harris TJR (1986) The cloning and nucleotide sequence of cDNA for an amplified glutamine synthetase gene from the Chinese hamster. Nucleic Acid Res. 14: 999–1008.

    Google Scholar 

  5. Hossell TE, Allen JC, Rowley AJ and Butler M (in press) The use of glutamine-free media for the growth of three cell lines in microcarrier. Develop. Biol. Standard.

  6. Iio M (1985) Influence of metabolites produced by cultured cell on cell proliferation in serum-free conditions. The Tissue Culture 11: 432–435.

    Google Scholar 

  7. Kovacevic Z (1971) The pathway of glutamine and glutamate oxidation in isolated mitochondria from mammalian cells. Biochem. J. 125: 757.

    Google Scholar 

  8. Kovacevic Z and Morris HP (1972) The role of glutamine in the oxidative metabolism of malignant cells. Cancer Res. 32: 326.

    Google Scholar 

  9. Kun E and Kearney EB (1981) Protein Metabolism: Ammonia. In: Bergmeyer HU (ed) Methods of Enzymatic Analysis (pp. 802–1806). Verlag Chemie International, Deerfield Beach, Florida.

    Google Scholar 

  10. Levintow L, Eagle H and Piez KA (1957a) The Role of Glutamine in Protein Biosynthesis in Tissue culture. J. Biol. Chem. 227: 929–941.

    Google Scholar 

  11. Lavietes BB, Regan DH and Demopoulos HB (1974) Glutamate oxidation of 6C3HED lymphoma: Effects of Lasparaginase on sensitive and resistant line. Proc. Natl. Acad. Sci. USA 71: 3993.

    Google Scholar 

  12. Levintow L (1957b) Evidence that glutamine is a precursor of asparagine in a human cell tissue culture. Science 126: 611–612.

    Google Scholar 

  13. Morris JE and Moscona AA (1970) Induction of glutamine synthetase in embryonic retina: its dependence on cell interactions. Science 167: 1736–1738.

    Google Scholar 

  14. Nagle SCJr (1968) Heat-stable chemically detined medium for growth of animal cell in suspension. Appl. Microbiol. 16: 53–55.

    Google Scholar 

  15. Niwa A, Fujiyoshi N, Yasumura Y and Mizuno H (1982) Establishment of a cell line from normal rat liver which grows continuously in protein-free chemically defined medium supplemented with no hormones or growth factors. Dokkyo J. Med. Sci. 9: 97–105.

    Google Scholar 

  16. Odland L, Wallin S and Walum E (1986) Lipid peroxidation and activities of tyrosine aminotransferase and glutamine synthetase in hepatoma and glioma cells grown in bovine colostrum-supplemented medium.In Vitro cellular and Develop. Biol. 22: 259–262.

    Google Scholar 

  17. Raivio KO and Seegmiller JE (1973) Adenine, hypoxanthine, and guanine metabolism in fibroblasts from normal individuals and from patients with hypoxanthine phosphori-bosyltransferase deficiency. Biochem. Biophys, Acta 299: 273–282.

    Google Scholar 

  18. Reizer LJ, Wice BM and Kennell D (1979) Evidence that glutamine, not sugar, is the major energy source for culture HeLa cells. J. Biol. Chem. 254: 2669–2676.

    Google Scholar 

  19. Sanders PG and Wilson RH (1984) Amplification and cloning of the Chinese hamster glutamine synthetase gene. EMBO J. 3: 65–71.

    Google Scholar 

  20. Sato S, Kawamura K and Fujiyoshi N (1983) Animal cell cultivation for production of Biological substances with a novel perfusion culture apparatus. J. Tissue Culture Methods 8: 167–171.

    Google Scholar 

  21. Thorndike J and Rief-Lehrer L (1971) A Sensitive Assay for Glutamultransferase. Enzyme 12: 235–241.

    Google Scholar 

  22. Yamane I and Arakawa K. Japanese Patent S. 47–670 (1972.1.10).

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Affiliations

  1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, 194, Machida-shi, Tokyo, Japan

    Shinji Hosoi, Hiroyuki Mioh, Chigusa Anzai, Seiji Sato & Nobuo Fujiyoshi

Authors
  1. Shinji Hosoi
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  2. Hiroyuki Mioh
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  3. Chigusa Anzai
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  4. Seiji Sato
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  5. Nobuo Fujiyoshi
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Hosoi, S., Mioh, H., Anzai, C. et al. Establishment of Namalva cell lines which grow continuously in glutamine-free medium. Cytotechnology 1, 151–158 (1988). https://doi.org/10.1007/BF00146816

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  • DOI: https://doi.org/10.1007/BF00146816

Key words

  • adaptation
  • ammonia
  • glutamine-free
  • glutamine synthetase
  • Namalva cells