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Cultivation of Trypanosoma brucei sspp. in semi-defined and defined media

Published online by Cambridge University Press:  06 April 2009

G. A. M. Cross
Affiliation:
Medical Research Council Biochemical Parasitology Unit, Molteno Institute, Downing Street, Cambridge CB2 3EE
J. C. Manning
Affiliation:
Medical Research Council Biochemical Parasitology Unit, Molteno Institute, Downing Street, Cambridge CB2 3EE

Extract

Semi-defined and defined media for the growth of culture forms of Trypanosoma brucei sspp. have been developed by enrichment of tissue culture medium 199 with additional vitamins, amino acids, salts and other compounds. The semi-defined medium contains an acid hydrolysate of casein: in the empirically devised defined medium the casein requirement has been circumvented by inclusion of additional vitamins and amino acids. Both media are very hypertonic. Control of pH was found to be particularly critical for growth. The optimum temperature for growth in the semidefined medium was between 25°C and 28°C, but cells would undergo one or two division cycles at 37°C.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1973

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References

REFERENCES

Bursell, E., (1970). Feeding digestion and excretion (in Glossina). In The African Trypanosomiases (ed. Mulligan, H. W.), pp. 305–16. London: George Allen and Unwin Ltd.Google Scholar
Chen, R. F. (1967). Removal of fatty acids from serum albumin by charcoal treatment. Journal of Biological Chemistry 242, 173–81.CrossRefGoogle ScholarPubMed
Cunningham, M. P. & Vickerman, K. (1962). Antigenic analysis in the Trypanosoma brucei group using the agglutination reaction. Transactions of the Royal Society of Tropical Medicine and Hygiene 56, 4859.CrossRefGoogle ScholarPubMed
Dixon, H. & Williamson, J. (1970). The lipid composition of blood and culture forms of Trypanosoma lewisi and Trypanosoma rhodesiense compared with that of their environment. Comparative Biochemistry and Physiology 33, 111–28.CrossRefGoogle ScholarPubMed
Dixon, H., Ginger, C. D. & Williamson, J. (1972). Trypanosome sterols and their metabolic origins. Comparative Biochemistry and Physiology 41B, 118.Google ScholarPubMed
Eagle, H. (1955 a). The minimum vitamin requirements of the L and HeLa cells in tissue culture, the production of specific vitamin deficiencies and their cure. Journal of Experimental Medicine 102, 595600.CrossRefGoogle Scholar
Eagle, H. (1955 b). The specific amino acid requirements of a mammalian cell (strain L) in tissue culture. Journal of Biological Chemistry 214, 839–52.CrossRefGoogle ScholarPubMed
Evans, D. A. & BROWN, R. C. (1971). Cyanide insensitive culture form of Trypanosoma brucei. Nature, London 230, 251–2.CrossRefGoogle ScholarPubMed
Good, N. E., Winget, G. D., Winter, W., Connolly, T. K., Izawa, S. & Singh, R. M. M. (1966). Hydrogen ion buffers for biological research. Biochemistry 5, 467–77.CrossRefGoogle ScholarPubMed
Haurowitz, F., & Hardin, R. L., (1954). Respiratory proteins. In The Proteins, vol. 2, part A, (ed. Neurath, H., and Bailey, K.), pp. 279344. New York: Academic Press.CrossRefGoogle Scholar
Holman, R. T., (1968). Essential fatty acid deficiency. In Progress in the chemistry of fats and other lipids, vol. 9, Polyunsaturated acids (ed. Holman, R. T.), part II, pp. 279348. Oxford: Pergamon Press.Google Scholar
Kidder, G. W. & Dutta, B. N. (1958). The growth and nutrition of Crithidia fasciculata. Journal of General Microbiology, 18, 621–38.CrossRefGoogle ScholarPubMed
Morgan, J. F., Morton, H. J. & Parker, R. C. (1950). Nutrition of animal cells in tissue culture. I. Initial studies on a synthetic medium. Proceedings of the Society for Experimental Medicine 73, 18.CrossRefGoogle ScholarPubMed
Newton, B. A., Cross, G. A. M., & Baker, J. R., (1973). Differentiation in Trypanosomatidae. In Microbial Differentiation, the Twenty-Third Symposium of the Society for General Microbiology (ed. Smith, J. E., and Ashworth, J. M.), pp. 339–73. Cambridge University Press.Google Scholar
Pittam, M. D. (1970). Medium for in vitro culture of Trypanosoma rhodesiense and T. brucei. Appendix to Dixo. Williamson (1970).Google Scholar
Rodbell, M. (1965). Modulation of lipolysis in adipose tissue by fatty acid concentration in fat cell. Annals of the New York Academy of Sciences 131, 302–14.CrossRefGoogle ScholarPubMed
Taylor, A. E. R., & Baker, J. R., (1968). The Cultivation of Parasites in vitro. Oxford: Blackwell Scientific Publications.Google Scholar
Tobie, E. J., Brand, T. von & Mehlman, B. (1950). Cultural and physiological observations on Trypanosoma rhodesiense and Trypanosoma gambiense. Journal of Parasitology 36, 4854.CrossRefGoogle ScholarPubMed
Trager, W. (1957). Nutrition of a hemoflagellate (Leishmania tarentolae) having an interchangeable requirement for choline or pyridoxal. Journal of Protozoology 4, 269–76.CrossRefGoogle Scholar
Vakertzi-Lemonias, C., Kidder, G. W. & Dewey, V. C. (1963). Ubiquinone in four genera of protozoa. Comparative Biochemistry and Physiology 8, 331–4.CrossRefGoogle Scholar
Vickerman, K., (1971). Morphological and physiological considerations of extracellular blood protozoa. In Ecology and Physiology of Parasites: A Symposium (ed. Fallis, A. M.), pp. 5891. Toronto: University Press.CrossRefGoogle Scholar
Weinman, D. (1960). Cultivation of the African sleeping sickness trypanosomes from the blood and cerebrospinal fluid of patients and suspects. Transactions of the Royal Society of Tropical Medicine and Hygiene 54, 180–90.CrossRefGoogle ScholarPubMed