Amoeba Culture Protocols for Research

The experiments were carried out using two amoeba species. N. fowleri type strain ATCC 30808™, from the American Type Culture Collection was used. The cells were axenically grown at 37°C in 2% bactocasitone medium supplemented with 10% of fetal bovine serum (FBS), 0.5 mg/mL of streptomycin sulphate and 0.3 μg/mL of penicillin G (Sigma Aldrich, Madrid, Spain). Clinical strain Acanthamoeba griffini genotype T3, obtained according to a previous study (González-Robles et al., 2014 (link)) was used. A. griffini trophozoites were grown axenically at 26°C in Peptone Yeast Glucose (PYG) medium (0.75% (w/v) proteose peptone, 0.75% (w/v) yeast extract, and 1.5% (w/v) glucose) containing 40 μg of gentamicin mL⁻¹ (Biowest, Nuaillé, France).