Tulahuen CL2 strain (nifurtimox-sensitive) was maintained on MRC-5 cells in minimal essential medium (MEM) supplemented with 20 mM L-Glutamine, 16.5 mM sodium hydrogen carbonate and FCS (5%) at 37 °C and 5% CO2. To determine in vitro Antitrypanosomal activity, 4 x 10 3 MRC-5 cells and 4 x 10 4 parasites were added to each well of test plate with compound.
After incubation at 37 °C for 7 days, parasite growth was assessed by adding of betagalactosidase substrate, chlorophenol red beta-D-galactopyranoside for 4 hours at 37 °C. The color reaction was read at 540 nm and absorbance values were expressed as a percentage of the blank controls. Nifurtimox was included as a reference drug.
After incubation at 37 °C for 7 days, parasite growth was assessed by adding of betagalactosidase substrate, chlorophenol red beta-D-galactopyranoside for 4 hours at 37 °C. The color reaction was read at 540 nm and absorbance values were expressed as a percentage of the blank controls. Nifurtimox was included as a reference drug.
