2.1.3. Amastigote and trypomastigote forms
Metacyclic trypomastigotes were used to infect Vero cells at 37ยบ C in humidified 95 % air, 5 % CO2 atmosphere in Gibco RPMI 1640 medium supplemented with 10% (v /v ) hiFBS (Contreraset al. , 1985; Pless-Petig et al. , 2012). After 5-7 days of incubation, culture-derived trypomastigotes were collected every 24 h by centrifuging the supernatant at 3000g for 5 min, and used to infect new Vero cells cultures and BALB/c mice.