Parasite Disease Group, Unit of Infection and Immunity, IBMC - Instituto de Biologia Molecular e Celular, Universidade do Porto, Portugal.
Centre de Recherche en Infectiologie du Centre de Recherche du CHUL, Québec, Canada.
Parasitology. 2014 Feb;141(2):192-205. doi: 10.1017/S0031182013001388. Epub 2013 Sep 6.
An ideal culture medium for Leishmania promastigotes should retain the basic characteristics of promastigotes found in sandflies (morphology and infectivity). Furthermore, the media should not create a bias in experimental settings, thus enabling the proper extrapolation of results. To assess this we studied several established media for promastigote growth. We analysed morphology, viability, cell cycle progression, metacyclic profile, capacity to differentiate into axenic amastigotes and infectivity. Furthermore, using a rational approach from the evaluated media we developed a simple serum-free medium (cRPMI). We report that parasites growing in different media present different biological characteristics and distinct in vitro and in vivo infectivities. The developed medium, cRPMI, proved to be a less expensive substitute for traditional serum-supplemented media for the in vitro maintenance of promastigotes. In fact, cRPMI is ideal for the maintenance of parasites in the laboratory, diminishing the expected loss of virulence over time typical of the parasite cultivation. Ultimately this report is a clear warning that the normalization of culture media should be a real concern in the field as media-specific phenomena are sufficient to induce biological bias with consequences in infectivity and general parasite biology.
用于利什曼原虫前鞭毛体的理想培养基应保留在沙蝇中发现的前鞭毛体的基本特征（形态和感染力）。此外，该培养基不应在实验环境中产生偏差，从而能够正确推断结果。为了评估这一点，我们研究了几种用于前鞭毛体生长的既定培养基。我们分析了形态、活力、细胞周期进程、循环期特征、分化为无细胞内阿米巴的能力和感染力。此外，我们从评估的培养基中采用合理的方法开发了一种简单的无血清培养基（cRPMI）。我们报告说，在不同培养基中生长的寄生虫具有不同的生物学特性，并且在体外和体内的感染力也不同。所开发的培养基 cRPMI 被证明是传统血清补充培养基的更便宜替代品，可用于体外维持前鞭毛体。事实上，cRPMI 非常适合在实验室中维持寄生虫，减少了随着时间的推移寄生虫培养中典型的毒力丧失。最终，本报告清楚地警告说，培养基的标准化应该是该领域真正关注的问题，因为培养基特异性现象足以引起生物学偏差，从而影响感染力和寄生虫的一般生物学。
