Skip to main content
Log in

Growth ofChlamydia psittaci strain menigopneumonitis in mouse L cells cultivated in a defined medium in spinner cultures

  • Published:
In Vitro Aims and scope Submit manuscript

Summary

L cells were grown in spinner cultures in a defined medium consisting of Waymouth medium MB752/1 (19) supplemented with 2 mg of fatty acid-free bovine serum albumin (BSA) per ml and 5 μg of oleate per ml (WO5 medium). Growth in WO5 medium was comparable to spinner L cell growth in two serum-containing media. The optimal concentration of oleate in the WO medium was 5 to 10 μg per ml. The use of 20 to 80 μg of oleate per ml of medium resulted in lower peak populations and earlier declines in viable cell counts. Cell death occurred rapidly in WO160 medium. Cell growth in WO medium containing 5 to 80 μg of oleate per ml was well above the level of growth observed when no oleate was present in the medium. Since the total lipid and fatty acid compositions of the BSA used in this study have been characterized by the authors, the WO medium may be considered a defined medium. L cells have been continuously maintained in spinner cultures in WO5 medium for over 50 passages with no major variation in the growth pattern. A 1000-fold increase inChlamydia psittaci strain meningopneumonitis, with a peak titer of 9.3×107 plaque-forming units per ml, was observed when the chlamydial agents were grown in spinner L cells in WO5 medium.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Moore, G. E., D. Mount, G. Tara, and N. Schwartz. 1963. Growth of human tumor cells in suspension cultures. Cancer Res. 23: 1735–1741.

    PubMed  CAS  Google Scholar 

  2. Fish, D. C., and J. P. Dobbs. 1971. Changes in enzyme specific activity and isoenzyme distribution patterns of L cells as a function of growth conditions. In Vitro 6: 441–450.

    PubMed  CAS  Google Scholar 

  3. Brown, B. L., and S. C. Nagle. 1965. Preservation of mammalian cells in a chemically defined medium and dimethylsulfoxide. Science 149: 1266–1267.

    Article  PubMed  CAS  Google Scholar 

  4. Bryant, J. C. 1966. Mammalian cells in chemically defined media in suspension culture. Ann. N. Y. Acad. Sci. 139: 143–161.

    Article  PubMed  CAS  Google Scholar 

  5. Bryant, J. C. 1970. Glucose and lactic acid trends in suspension cultures of two established mammalian cell strains in chemically defined media. Biotechnol. Bioeng. 12: 429–464.

    Article  PubMed  CAS  Google Scholar 

  6. Higuchi, K. 1970. An improved chemically defined culture medium for strain L mouse cells based on growth responses to graded levels of nutrients including iron and zinc ions. J. Cell Physiol. 75: 65–72.

    Article  PubMed  CAS  Google Scholar 

  7. Lengle, E., and R. P. Geyer. 1972. Comparison of cellular lipids of serum-free strain L mouse fibroblasts. Biochim. Biophys. Acta 260: 608–616.

    PubMed  CAS  Google Scholar 

  8. Merchant, D. J., J. S. Walker, and F. R. Parker. 1966. Some effects of cultural conditions and media upon selection of variants within an animal cell line. In Vitro 2: 127.

    Google Scholar 

  9. Nagle, S. C. 1968. Heat-stable chemically defined medium for growth of animal cells in suspension. Appl. Microbiol. 16: 53–55.

    PubMed  Google Scholar 

  10. Weirether, F. J., J. S. Walker, and R. E. Lincoln. 1968. A precise method for replicating suspension cultures of mammalian cells. Appl. Microbiol. 16: 841–844.

    PubMed  CAS  Google Scholar 

  11. Quash, G., E. Delain, and J. Huppert. 1971. Effect of antipolyamine antibodies on mammalian cells in tissue culture. Exp. Cell Res. 66: 426–432.

    Article  PubMed  CAS  Google Scholar 

  12. Tribby, I. I. E., and J. W. Moulder. 1966. Availability of bases and nucleosides as precursors of nucleic acids in L cells and in the agent of meningopneumonitis. J. Bacteriol. 91: 2362–2367.

    PubMed  CAS  Google Scholar 

  13. Schechter, E. M. 1966. Synthesis of nucleic acid and protein in L cells infected with the agent of meninopneumonitis. J. Bacteriol. 91: 2069–2080.

    PubMed  CAS  Google Scholar 

  14. Hull, R. N., W. R. Cherry, and O. J. Tritch. 1962. Growth characteristics of monkey kidney cell strains LLC-MK1, LLC-MK2, and LLC-MK2(NCTC-3196) and their utility in virus research. J. Exp. Med. 115 903–918.

    Article  PubMed  CAS  Google Scholar 

  15. Jenkin, H. M., and L. E. Anderson. 1970. The effect of oleic acid on the growth of monkey kidney cells (LLC-MK2). Exp. Cell Res. 59: 6–10.

    Article  PubMed  CAS  Google Scholar 

  16. Makino, S., H. M. Jenkin, H. M. Yu, and D. Townsend. 1970. Lipid composition ofChlamydia psittaci grown in monkey kidney cells in defined medium. J. Bacteriol. 103: 62–70.

    PubMed  CAS  Google Scholar 

  17. Earle, W. R. 1943. Production of malignancyin vitro. IV. The mouse fibroblast cultures and changes seen in the living cell. J. Nat. Cancer Inst. 4: 165–212.

    CAS  Google Scholar 

  18. Sanford, K. K., W. R. Earle, and G. D. Likely. 1948. The growthin vitro of single isolated cells. J. Nat. Cancer Inst. 9: 229–246.

    Google Scholar 

  19. Waymouth, C. 1959. Rapid proliferation of sublines of NCTC clone 929 (strain L) mouse cells in a simple chemically defined medium (MB 752/1). J. Nat. Cancer Inst. 22: 1003–1017.

    PubMed  CAS  Google Scholar 

  20. Jenkin, H. M., L. E. Anderson, R. T. Holman, I. A. Ismail, and F. D. Gunstone. 1969. Effect of isomericcis-octadecenoic acids on the growth ofLeptospira interrogans serotypepactoc. J. Bacteriol. 98: 1026–1029.

    PubMed  CAS  Google Scholar 

  21. Morgan, J. F., H. J. Morton, and R. C. Parker. 1950. Nutrition of animal cells in tissue culture. I. Initial studies on a synthetic medium. Proc. Soc. Exp. Biol. Med. 73: 1–8.

    PubMed  CAS  Google Scholar 

  22. McLimans, W. F., F. E. Giardinello, E. V. Davis, C. J. Kucera, and G. W. Rake. 1957. Submerged culture of mammalian cells: the five liter fermentor. J. Bacteriol. 74: 768–774.

    PubMed  CAS  Google Scholar 

  23. Francis, T. J., and T. P. Magill. 1938. Umdentified virus causing acute meningitis and pneumonitis in experimental animals. J. Exp. Med. 68: 147–160.

    Article  Google Scholar 

  24. Jenkin, H. M. 1966. Continuous passage of agents of trachoma in cell culture. I. Characteristics of TW-3 and Bour strains of trachoma cultivated in serial passage in HeLa 229 cells. J. Infect. Dis. 116: 390–399.

    PubMed  CAS  Google Scholar 

  25. Alexander, J. J. 1969. Effect of infection with the meningopneumonitis agent on deoxyribonucleic acid and protein by its L-cell host. J. Bacteriol. 97: 653–657.

    Article  PubMed  CAS  Google Scholar 

  26. Banks, J., B. Eddie, J. Schachter, and K. F. Meyer. 1970. Plaque formation byChlamydia in L cells. Infect. Immun. 1: 259–262.

    PubMed  Google Scholar 

  27. Eagle, H. 1959. Amino acid metabolism in mammalian cell cultures. Science 130: 432–437.

    Article  PubMed  CAS  Google Scholar 

  28. Hanks, J. H., and R. E. Wallace. 1949. Relation of oxygen and temperature in the preservation of tissues by refrigeration. Proc. Soc. Exp. Biol. Med. 71: 196–200.

    Google Scholar 

  29. Jenkin, H. M., S. Makino, D. Townsend, M. C. Riera, and A. L. Barron. 1970. Lipid composition of the hemagglutinating active fraction obtained from chick embryos infected withChlamydia psittaci 6BC. Infect. Immun. 2: 316–319.

    PubMed  CAS  Google Scholar 

  30. Chen, R. F. 1967. Removal of fatty acids from serum albumin by characoal treatment. J. Biol. Chem. 242: 173–181.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

This investigation was supported by Public Health Service Research Grant HE 08214 from the Program Projects Branch, Extramural Programs, National Heart and Lung Institute; The World Health Organization; and The Hormel Foundation.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Morrison, S.J., Jenkin, H.M. Growth ofChlamydia psittaci strain menigopneumonitis in mouse L cells cultivated in a defined medium in spinner cultures. In Vitro 8, 94–100 (1972). https://doi.org/10.1007/BF02615966

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02615966

Key words

Navigation