Yabu Y, Koide T, Ohta N, Nose M, Ogihara Y
Department of Medical Zoology, Nagoya City University Medical School, Nagoya, Japan.
Southeast Asian J Trop Med Public Health. 1998 Sep;29(3):591-5.
An effective axenic culture system for bloodstream forms of Trypanosoma brucei brucei GUT at 3.1 containing a low concentration of serum is described. Bloodstream forms routinely maintained in Iscove's modification of Dulbecco's medium supplemented with 100 microM hypoxanthine, 30 microM thymidine, 40 microM adenosine, 1 mM sodium pyruvate, 50 microM L-glutamine, 100 microM 2-mercaptoethanol and 20% FBS for more than one year were grown in the same medium supplemented with 5% FBS without reducing their growth rate. Then culture adapted trypanosomes in the culture medium containing 5% FBS were transferred into the modified medium supplemented with 0.5% FBS. For the constant growth of bloodstream forms in the medium containing 0.5% FBS, the culture medium was further supplemented with 200 microM L-alanine, 100 microM glycine, 10 microM L-oruithine hydrochloride and 10 microM L-citrullin. The trypanosomes propagated in this culture system for one year retained their infectivity for mice. This culture system was also shown to be useful for cloning of T.b. brucei GUT at 3.1 which is important for separation of mutants.
本文描述了一种有效的无共生培养系统，用于培养布氏布氏锥虫（Trypanosoma brucei brucei）GUTat 3.1的血流形式，该系统含有低浓度血清。常规在添加了100微摩尔次黄嘌呤、30微摩尔胸苷、40微摩尔腺苷、1毫摩尔丙酮酸钠、50微摩尔L-谷氨酰胺、100微摩尔2-巯基乙醇和20%胎牛血清的Iscove改良杜氏培养基中培养一年以上的血流形式，在添加5%胎牛血清的相同培养基中生长，且生长速率未降低。然后将在含有5%胎牛血清的培养基中适应培养的锥虫转移到添加了0.5%胎牛血清的改良培养基中。为了使含有0.5%胎牛血清的培养基中的血流形式持续生长，培养基进一步添加了200微摩尔L-丙氨酸、100微摩尔甘氨酸、10微摩尔L-鸟氨酸盐酸盐和10微摩尔L-瓜氨酸。在该培养系统中繁殖一年的锥虫对小鼠仍具有感染性。该培养系统还被证明可用于克隆布氏布氏锥虫GUTat 3.1，这对于分离突变体很重要。
