半胱氨酸消除了体外培养布氏锥虫血流形式对饲养细胞的需求。
Cysteine eliminates the feeder cell requirement for cultivation of Trypanosoma brucei bloodstream forms in vitro.
作者信息
Duszenko M, Ferguson M A, Lamont G S, Rifkin M R, Cross G A
出版信息
J Exp Med. 1985 Oct 1;162(4):1256-63. doi: 10.1084/jem.162.4.1256.
Abstract
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 greater than 24 mg/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 Km for cysteine uptake was 4 X 10(-4) M. Cystine supported axenic growth if low concentrations of 2-mercaptoethanol were added at regular intervals.
摘要
在以往所有研究中，布氏锥虫的血流型仅在哺乳动物成纤维细胞饲养层的支持下才能在体外培养。我们通过定期添加适当浓度的L-半胱氨酸，在无饲养层条件下培养了布氏锥虫的血流型。最佳半胱氨酸浓度取决于细胞密度，且接近生理血清水平。当浓度高于24毫克/升（2×10⁻⁴摩尔/升）时，半胱氨酸对浓度为3×10⁷/毫升的锥虫具有急性毒性。添加丙酮酸或过氧化氢酶可防止毒性，它们能中和半胱氨酸自氧化产生的过氧化氢。在使用[³⁵S]半胱氨酸和[³⁵S]胱氨酸的摄取研究中，布氏锥虫仅能有效摄取半胱氨酸。半胱氨酸摄取的米氏常数为4×10⁻⁴摩尔/升。如果定期添加低浓度的2-巯基乙醇，胱氨酸可支持无饲养层培养。
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