Laboratório de Imunopatologia, Núcleo de Pesquisas em Ciências Biológicas/NUPEB, Universidade Federal de Ouro Preto, Morro do Cruzeiro, Ouro Preto, Minas Gerais, 35400-000, Brazil.
Laboratório de Morfopatologia, Núcleo de Pesquisas em Ciências Biológicas/NUPEB, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil.
Parasitol Res. 2020 Dec;119(12):4185-4195. doi: 10.1007/s00436-020-06893-z. Epub 2020 Oct 9.
Leishmania spp. parasites have a complex biological cycle presenting basically two different morphological stages, the amastigote and promastigote forms. In vitro cultivation allows a more complete study of the biological aspects of these parasites, indicating better conditions for infection, immunoassay tests, drug evaluations, and vaccines. Thus, we evaluated the three most used culture media for Leishmania spp., Grace's insect cell culture medium (Grace's), liver infusion tryptose (LIT), and Schneider's insect medium (Schneider's), without supplementation or supplemented with fetal calf serum (FCS) and bovine serum albumin (Albumin) to evaluate the growth, viability, and infectivity of the L. infantum promastigotes. It was observed that promastigote forms have a better growth in LIT and Schneider's with or without FCS when compared to that in Grace's. The supplementation with albumin promoted greater viability of the parasites independent of the medium. For in vitro infection of J774.A1 macrophages using light microscopy and flow cytometry analyses, FCS-supplemented LIT and Grace's promoted higher percentage of infected macrophages and parasite load compared with Schneider's media. Taken together, our results demonstrated that the supplementation of LIT culture medium with FCS is the most suitable strategy to cultivate Leishmania infantum parasites enabling the maintenance of growth and infective parasites for research uses.
利什曼原虫寄生虫具有复杂的生物学循环，主要呈现两种不同的形态阶段，即无鞭毛体和前鞭毛体。体外培养可更全面地研究这些寄生虫的生物学特性，为感染、免疫检测试验、药物评估和疫苗接种提供更好的条件。因此，我们评估了三种最常用于培养利什曼原虫的培养基，即格雷斯昆虫细胞培养基（Grace's）、肝浸液色氨酸（LIT）和施耐德昆虫培养基（Schneider's），不添加或添加胎牛血清（FCS）和牛血清白蛋白（Albumin），以评估婴儿利什曼原虫前鞭毛体的生长、活力和感染力。结果表明，与 Grace's 相比，前鞭毛体在 LIT 和 Schneider's 中无论是否添加 FCS 都有更好的生长。无论培养基如何，白蛋白的添加都能提高寄生虫的活力。使用荧光显微镜和流式细胞术分析 J774.A1 巨噬细胞的体外感染，结果表明，与 Schneider's 培养基相比，添加 FCS 的 LIT 和 Grace's 培养基促进了更高比例的感染巨噬细胞和寄生虫载量。综上所述，我们的研究结果表明，在 LIT 培养基中添加 FCS 是培养婴儿利什曼原虫寄生虫最适宜的策略，能够维持生长和感染性寄生虫用于研究用途。
