Entamoeba histolytica: cyst-like structures in vitro induction

The cyst of Entamoeba histolytica is responsible for amebiasis infection. However, no axenic in vitro system exists that promotes mass encystation for studying this process of this human-infecting parasite. Cyst-like structures of E. histolytica obtained in this work were induced using TYI-S-33 media in combination with enterobacterias Escherichia coli and Enterococcus faecalis conditioned media, high CO2 tension and histamine. Cyst-like structures showed the same characteristics of a typical E. histolytica cyst: aggregation, resistance to 0.15% sarcosyl for 10 min, high signal of fluorescence under UV light when stained with 10% calcofluor M2r and the surface topology showed a wrinkled wall. In addition these structures are multinucleated with condensed chromatin attached to nuclear membrane, contain big vacuoles and ribonucleoproteic helices in the cytoplasm and also present a thin cell wall. Last all characteristics are all the same as a typical of E. histolytica cyst.