Trypanosoma cruzi: Selective Isolation of Pure Trypomastigotes from Cultured Muscle Cells

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Abstract
Trypomastigote or trypomastigote-amastigote populations of T. cruzi (Y strain, MERC 2C) entierely free of epimastigotes were obtained from infected muscle cell cultures and separated from host-cell debris by passage through a DEAE-Cellulose column. Approximately 75% of the parasites were recovered and mouse infectivity titrations with postcolumn trypomastigotes showed only a slight reduction in infectivity compared to starting material. Light microscopy and EM examination of material showed that a high degree of purity was achieved by the column procedure. No host-cell debris could be identified in the eluate and parasites were morphologically intact. Enumeration of trypomastigotes and amastigotes in mixed populations before and after column purification showed that there was no preferential loss of either stage and both had the same residence time. This procedure may be used to obtain clean, minimally altered, parasite material free of the invertebrate epimastigote stage and host-cell debris for studies which are sensitive to these contaminants.