2.1.2. Transformation to metacyclic forms
Metacyclic trypomastigotes were induced by culturing a 7-day-old culture of epimastigote forms at 28 °C in Gibco Grace’s Insect Medium supplemented with 10% (v/v) hiFBS (Isola et al. , 1986). Subsequently, parasites were incubated at a density of 5 × 108 mL−1 for 2 h at 28 °C in TAU medium (190 mM NaCl, 17 mM KCl, 2 mM CaCl2, 2 mM MgCl2, 8 mM phosphate buffer, pH 6.0), and later at a density of 5 × 106 mL−1 for 4 days at 28 °C in TAU3AAG medium (TAU supplemented with 50 mM L-sodium glutamate, 2 mM L-sodium aspartate, 10 mM L-proline, 10 mM D-glucose) (Cardoso and Soares, 2010).