Cysteine eliminates the feeder cell requirement for cultivation of Trypanosoma brucei bloodstream forms in vitro

TY - JOUR

T1 - Cysteine eliminates the feeder cell requirement for cultivation of Trypanosoma brucei bloodstream forms in vitro

AU - Duszenko, Michael

AU - Ferguson, Michael A. J.

AU - Lamont, Gretel S.

AU - Rifkin, Mary R.

AU - Cross, George A.

PY - 1985/10/1

Y1 - 1985/10/1

N2 - In all previous studies, bloodstream forms of Trypanosoma brucei could be grown in vitro only when supported by a feeder layer of mammalian fibroblasts. We have axenically cultivated bloodstream T. brucei by adding L-cysteine at regular intervals and appropriate concentrations. The optimum cysteine concentration depends on cell density and is close to physiological serum levels. At concentrations >24mg/liter (2 x 10(-4) M), cysteine was acutely toxic to trypanosome concentrations of 3 x 10(7)/ml. Toxicity was prevented by addition of pyruvate or catalase, which neutralize H2O2 produced by cysteine autoxidation. In uptake studies using [35S]cysteine and [35S]cystine, T. brucei efficiently incorporated only cysteine. The K(m) for cysteine uptake was 4 x 10(-4) M. Cystine supported axenic growth if low concentrations of 2-mercaptoethanol were added at regular intervals.

AB - In all previous studies, bloodstream forms of Trypanosoma brucei could be grown in vitro only when supported by a feeder layer of mammalian fibroblasts. We have axenically cultivated bloodstream T. brucei by adding L-cysteine at regular intervals and appropriate concentrations. The optimum cysteine concentration depends on cell density and is close to physiological serum levels. At concentrations >24mg/liter (2 x 10(-4) M), cysteine was acutely toxic to trypanosome concentrations of 3 x 10(7)/ml. Toxicity was prevented by addition of pyruvate or catalase, which neutralize H2O2 produced by cysteine autoxidation. In uptake studies using [35S]cysteine and [35S]cystine, T. brucei efficiently incorporated only cysteine. The K(m) for cysteine uptake was 4 x 10(-4) M. Cystine supported axenic growth if low concentrations of 2-mercaptoethanol were added at regular intervals.

UR - https://www.scopus.com/pages/publications/0022410467

U2 - 10.1084/jem.162.4.1256

DO - 10.1084/jem.162.4.1256

M3 - Article

AN - SCOPUS:0022410467

SN - 0022-1007

VL - 162

SP - 1256

EP - 1263

JO - Journal of Experimental Medicine

JF - Journal of Experimental Medicine

IS - 4

ER -