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. 1980 Oct;77(10):5993–5997. doi: 10.1073/pnas.77.10.5993

Growth of T-lymphoma cells in serum-free medium: lack of involvement of the cyclic AMP pathway in long-term cultures.

F J Darfler, H Murakami, P A Insel
PMCID: PMC350199  PMID: 6255474

Abstract

We have developed a serum-free, chemically defined growth medium containing casein, insulin, transferrin, testosterone, and linoleic acid in Dulbecco's modified Eagle's medium/Ham's F12 medium, 1:1 (vol/vol), for growing murine T lymphomas. This medium supports the growth in suspension of all murine T lymphomas tested, including S49, WEHI 7, EL4, BW5147, and R1.1. Growth of these cell lines was maintained indefinitely with doubling times approaching those of cells grown in 10% (vol/vol) horse serum. This medium also supports the growth of several of the S49 variants of the beta-adrenergic receptor/adenylate cyclase/cyclic AMP/protein kinase pathway, suggeting little or no involvement of this pathway in the routine growth of S49 cells or in the mechanism of action of the factors in this defined medium. This serum-free medium should prove useful for studies of a variety of metabolic pathways and of differentiated functions of T-lymphoma cells.

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Selected References

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  1. Barnes D., Sato G. Methods for growth of cultured cells in serum-free medium. Anal Biochem. 1980 Mar 1;102(2):255–270. doi: 10.1016/0003-2697(80)90151-7. [DOI] [PubMed] [Google Scholar]
  2. Bottenstein J., Hayashi I., Hutchings S., Masui H., Mather J., McClure D. B., Ohasa S., Rizzino A., Sato G., Serrero G. The growth of cells in serum-free hormone-supplemented media. Methods Enzymol. 1979;58:94–109. doi: 10.1016/s0076-6879(79)58127-0. [DOI] [PubMed] [Google Scholar]
  3. Chlapowski F. J., Kelly L. A., Butcher R. W. Cyclic nucleotides in cultured cells. Adv Cyclic Nucleotide Res. 1975;6:245–338. [PubMed] [Google Scholar]
  4. Coffino P., Bourne H. R., Friedrich U., Hochman J., Insel P. A., Lemaire I., Melmon K. L., Tomkins G. M. Molecular mechanisms of cyclic AMP action: a genetic approach. Recent Prog Horm Res. 1976;32:669–684. doi: 10.1016/b978-0-12-571132-6.50037-3. [DOI] [PubMed] [Google Scholar]
  5. Coffino P., Bourne H. R., Insel P. A., Melmon K. L., Johnson G., Vigne J. Studies of cyclic AMP action using mutant tissue culture cells. In Vitro. 1978 Jan;14(1):140–145. doi: 10.1007/BF02618180. [DOI] [PubMed] [Google Scholar]
  6. Coffino P., Bourne H. R., Tomkins G. M. Somatic genetic analysis of cyclic AMP action: selection of unresponsive mutants. J Cell Physiol. 1975 Jun;85(3):603–610. doi: 10.1002/jcp.1040850312. [DOI] [PubMed] [Google Scholar]
  7. Coffino P., Gray J. W., Tomkins G. M. Cyclic AMP, a nonessential regulator of the cell cycle. Proc Natl Acad Sci U S A. 1975 Mar;72(3):878–882. doi: 10.1073/pnas.72.3.878. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen A., Ullman B., Martin D. W., Jr Characterization of a mutant mouse lymphoma cell with deficient transport of purine and pyrimidine nucleosides. J Biol Chem. 1979 Jan 10;254(1):112–116. [PubMed] [Google Scholar]
  9. Friedman D. L., Johnson R. A., Zeilig C. E. The role of cyclic nucleotides in the cell cycle. Adv Cyclic Nucleotide Res. 1976;7:69–114. [PubMed] [Google Scholar]
  10. Haga T., Ross E. M., Anderson H. J., Gilman A. G. Adenylate cyclase permanently uncoupled from hormone receptors in a novel variant of S49 mouse lymphoma cells. Proc Natl Acad Sci U S A. 1977 May;74(5):2016–2020. doi: 10.1073/pnas.74.5.2016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Harris A. W., Bankhurst A. D., Mason S., Warner N. L. Differentiated functions expressed by cultured mouse lymphoma cells. II. Theta antigen, surface immunoglobulin and a receptor for antibody on cells of a thymoma cell line. J Immunol. 1973 Feb;110(2):431–438. [PubMed] [Google Scholar]
  12. Honma Y., Kasukabe T., Okabe J., Hozumi M. Replacement of serum by insulin, transferrin, albumin, phosphatidyl choline, cholesterol, and some trace elements in cultures of mouse myeloid leukemia cells sensitive to inducers of differentiation. Exp Cell Res. 1979 Dec;124(2):421–428. doi: 10.1016/0014-4827(79)90217-9. [DOI] [PubMed] [Google Scholar]
  13. Horibata K., Harris A. W. Mouse myelomas and lymphomas in culture. Exp Cell Res. 1970 Apr;60(1):61–77. doi: 10.1016/0014-4827(70)90489-1. [DOI] [PubMed] [Google Scholar]
  14. Insel P. A., Bourne H. R., Coffino P., Tomkins G. M. Cyclic AMP-dependent protein kinase: pivotal role in regulation of enzyme induction and growth. Science. 1975 Nov 28;190(4217):896–898. doi: 10.1126/science.171770. [DOI] [PubMed] [Google Scholar]
  15. Iscove N. N., Melchers F. Complete replacement of serum by albumin, transferrin, and soybean lipid in cultures of lipopolysaccharide-reactive B lymphocytes. J Exp Med. 1978 Mar 1;147(3):923–933. doi: 10.1084/jem.147.3.923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Johnson G. L., Bourne H. R., Gleason M. K., Coffino P., Insel P. A., Melmon K. L. Isolation and characterization of S49 lymphoma cells deficient in beta-adrenergic receptors: relation of receptor number to activation of adenylate cyclase. Mol Pharmacol. 1979 Jan;15(1):16–27. [PubMed] [Google Scholar]
  17. Levinson B. B., Ullman B., Martin D. W., Jr Pyrimidine pathway variants of cultured mouse lymphoma cells with altered levels of both orotate phosphoribosyltransferase and orotidylate decarboxylase. J Biol Chem. 1979 Jun 10;254(11):4396–4401. [PubMed] [Google Scholar]
  18. Old L. J., Boyse E. A., Stockert E. The G (Gross) leukemia antigen. Cancer Res. 1965 Jul;25(6):813–819. [PubMed] [Google Scholar]
  19. Ralph P. Retention of lymphocyte characteristics by myelomas and theta + -lymphomas: sensitivity to cortisol and phytohemagglutinin. J Immunol. 1973 Jun;110(6):1470–1475. [PubMed] [Google Scholar]
  20. Raschke W. C. Plasmacytomas, lymphomas and hybridomas: their contribution to immunology and molecular biology. Biochim Biophys Acta. 1980 Mar 12;605(1):113–145. doi: 10.1016/0304-419x(80)90023-2. [DOI] [PubMed] [Google Scholar]
  21. Roubinian J. R., Talal N., Greenspan J. S., Goodman J. R., Siiteri P. K. Effect of castration and sex hormone treatment on survival, anti-nucleic acid antibodies, and glomerulonephritis in NZB/NZW F1 mice. J Exp Med. 1978 Jun 1;147(6):1568–1583. doi: 10.1084/jem.147.6.1568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wolfe R. A., Wu R., Sato G. H. Epidermal growth factor-induced down-regulation of receptor does not occur in HeLa cells grown in defined medium. Proc Natl Acad Sci U S A. 1980 May;77(5):2735–2739. doi: 10.1073/pnas.77.5.2735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Yamamoto K. R., Gehring U., Stampfer M. R., Sibley C. H. Genetic approaches to steroid hormone action. Recent Prog Horm Res. 1976;32:3–32. doi: 10.1016/b978-0-12-571132-6.50008-7. [DOI] [PubMed] [Google Scholar]

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