Liver infusion tryptose (LIT): the best choice for growth, viability, and infectivity of Leishmania infantum parasites

肝灌注胰蛋白胨 (LIT):利什曼原虫生长、活力和感染力的最佳选择

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作者:Ana Flávia Pereira Costa, Rory Cristiane Fortes de Brito, Lívia Mendes Carvalho, Jamille Mirelle de Oliveira Cardoso, Paula Melo de Abreu Vieira, Alexandre Barbosa Reis, Rodrigo Dian de Oliveira Aguiar-Soares, Bruno Mendes Roatt

Abstract

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.

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