Development of a three-dimensional migration assay for study of Trypanosoma cruzi-mammalian cell interaction.

RODRIGUEZ, ME; BÚA, J; CAMPO, VA; LEVY, GV; SÁNCHEZ, DO; TEKIEL V

Caxambú

Congreso; XXXIII Reunião Anual da Sociedade Brasileira de Protozoologia/XLIV Reunião Anual sobre Pesquisa Básica em Doença de Chagas; 2017

Sociedade Brasileira de Protozoologia

Development of a three-dimensional migration assay for study of Trypanosoma cruzi-mammalian cell interactionRODRÍGUEZ, M.E., BÚA J., CAMPO V.A., LEVY, G., SANCHEZ, D.O.. TEKIEL, V. Three-dimensional (3D) cultures are physiologically relevant because mimic the microarchitecture of tissues and provide an environment similar to the encountered in natural infections, bridging the gap between traditional monolayer cultures (2D) and animal models. In this work, we combined the 3D culture technology with host-pathogen interaction, by studying transmigration of T. cruzi trypomastigotes of two representative strains (CL Brener DTU VI; SylvioX10 DTU I) into 3D spheroids. Spheroids of HeLa cells expressing Actin-RFP were cultured with CFSE-labelled trypomastigotes and transmigration into spheroids was analyzed at 24 h post infection. Images obtained by 3D-reconstruction from confocal microscopy stacks showed that CL Brener was more migratory than SylvioX10, which was retained at the surface of spheroids (>40 µm vs