Culturing Naegleria fowleri Trophozoites

Trophozoites of two clinic isolates of Naegleria fowleri from Costa Rica (accession numbers MT090627 and MT210902) (Retana Moreira et al., 2020b (link)) were cultured in 75 cm² Nunc EasYFlask cell culture flasks (Thermo Fisher Scientific, Waltham, Massachusetts, United States) with 2% casein hydrolysate (Sigma Aldrich, Missouri, United States) culture medium, supplemented with 10% inactivated fetal bovine serum (Gibco, GranIsland, New York, United States) and antibiotics (penicillin/streptomycin). The flasks were incubated at 37°C, with daily observation of the cultures under an inverted microscope. For each flask, the culture medium was replaced, at least, every 2 days.