Growth ofChlamydia psittaci strain menigopneumonitis in mouse L cells cultivated in a defined medium in spinner cultures
- 38 Downloads
- 13 Citations
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.
Key words
defined medium L cells spinner culture Chlamydia psittaci strain meningopneumonitis Waymouth oleate mediumPreview
Unable to display preview. Download preview PDF.
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.PubMedGoogle 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.PubMedGoogle 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.PubMedCrossRefGoogle Scholar
- 4.Bryant, J. C. 1966. Mammalian cells in chemically defined media in suspension culture. Ann. N. Y. Acad. Sci. 139: 143–161.PubMedCrossRefGoogle 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.PubMedCrossRefGoogle 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.PubMedCrossRefGoogle 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.PubMedGoogle 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.PubMedGoogle 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.PubMedGoogle 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.PubMedCrossRefGoogle 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.PubMedGoogle 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.PubMedGoogle 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.PubMedCrossRefGoogle 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.PubMedCrossRefGoogle 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.PubMedGoogle 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.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.PubMedGoogle 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.PubMedGoogle 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.PubMedGoogle 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.PubMedGoogle 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.CrossRefGoogle 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.PubMedGoogle 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.PubMedCrossRefGoogle Scholar
- 26.Banks, J., B. Eddie, J. Schachter, and K. F. Meyer. 1970. Plaque formation byChlamydia in L cells. Infect. Immun. 1: 259–262.PubMedGoogle Scholar
- 27.Eagle, H. 1959. Amino acid metabolism in mammalian cell cultures. Science 130: 432–437.PubMedCrossRefGoogle 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.PubMedGoogle Scholar
- 30.Chen, R. F. 1967. Removal of fatty acids from serum albumin by characoal treatment. J. Biol. Chem. 242: 173–181.PubMedGoogle Scholar