Liver infusion tryptose (LIT): the best choice for growth, viability, and infectivity of Leishmania infantum parasites
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PROTOZOOLOGY - ORIGINAL PAPER
Liver infusion tryptose (LIT): the best choice for growth, viability, and infectivity of Leishmania infantum parasites Ana Flávia Pereira Costa 1 & Rory Cristiane Fortes de Brito 1 & Lívia Mendes Carvalho 1 & Jamille Mirelle de Oliveira Cardoso 1 & Paula Melo de Abreu Vieira 2,3 & Alexandre Barbosa Reis 1,4 & Rodrigo Dian de Oliveira Aguiar-Soares 1 & Bruno Mendes Roatt 1,3,4 Received: 13 May 2020 / Accepted: 16 September 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
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. Keywords Leishmania infantum . In vitro cultivation . Liver infusion tryptose . Grace’s insect cell culture medium . Schneider’s insect medium
Introduction Section Editor: Nawal Hijjawi Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00436-020-06893-z) contains supplementary material, which is available to authorized users. * Bruno Mendes Roatt [email protected]; [email protected] 1
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
2
Laboratório de Morfopatologia, Núcleo de Pesquisas em Ciências Biológicas/NUPEB, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil
3
Departamento de Ciências Biológicas, Insituto de Ciências Exatas e Biológicas, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil
4
Instituto Nacional de Ciência e Tecnologia em Doenças Tropicais, INCT-DT, Salvador, Brazil
Leishmania spp. parasites have a life cycle involving both a mammalia
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