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Hesse, F; Selzer, PM; Mühlstädt, K; Duszenko, M.
(1995): A novel cultivation technique for long-term maintenance of bloodstream form trypanosomes in vitro.
Mol Biochem Parasitol. 1995; 70(1-2):157-166

Abstract:: We used-an axenic cultivation system to grow African trypanosomes in vitro. Long-term cultivation for more than 60 days has been achieved by replacing the culture medium at regular intervals between 6 and 48 h. In contrast to a control culture without medium replacement, increasing amounts of maximum cell concentrations have been obtained, ranging from 5 x 10(6) to 2 X 10(7) trypanosomes ml(-1), whereas the generation doubling time remained constant (about 6 h). Higher cell concentrations have only been obtained by total medium replacement; neither addition of fresh medium nor serum led to a higher cell yield, suggesting that a trypanosome-derived factor or metabolite accumulated in the medium rather than medium was depleted of an essential nutrient. Most interestingly, however, successive waves have been obtained which eventually led to a damped oscillation curve with a constant high population density after about 40 days of cultivation. Cultures were started with a homogeneous population of the long-slender form. As judged by light microscopy, cells showed a stumpy morphology during the declining phase and became slender again in the following growth phase. At later time points, when cells remained in a stationary phase at high population density, many different morphological stages have been observed, similar to those described by early authors as intermediate forms [Ormerod, W.E. (1979) In: Biology of the Kinetoplastida, Vol. 2, pp. 340-393], although many dividing forms are still present at that time. In contrast, identically treated procyclic cultures were unable to produce cyclic growth waves. Based on these results, a novel concept considering a possible differentiation mechanism is discussed.
Authors BOKU Wien:: Hesse Friedemann
Find related publications in this database (using NML MeSH Indexing): Animals -; Blood Physiological Phenomena -; Culture Media, Conditioned - chemistry; Growth Inhibitors - isolation & purification; Mice -; Mice, Inbred Strains -; Molecular Weight -; Parasitemia - parasitology; Parasitology - methods; Rats -; Rats, Sprague-Dawley -; Trypanosoma brucei brucei - growth & development; Trypanosomiasis, African - blood
Find related publications in this database (Keywords): TRYPANOSOMA BRUCEI; AXENIC CULTIVATION; DIFFERENTIATION