Small-scale in vitro culture and purification of Plasmodium berghei for transfection experiment

The standard protocol for genetic modification of the rodent malaria parasite Plasmodium berghei requires infected blood from one or more laboratory mice, followed by large-scale in vitro parasite culture and purification of mature schizonts. Here, protocols are described for small-scale in vitro culture from 20 μL of mouse tail blood and purification of mature P. berghei schizonts sufficient for a single transfection experiment. All procedures are performed in 1.5-mL microcentrifuge tubes. We confirmed that transgenic parasites could be obtained using schizonts prepared by this protocol. This small-scale protocol provides significant advantages, namely reduction of parasite sample, laboratory consumables and mice for transfection experiments.

Protocols for small-scale in vitro culture from 20 μL of mouse tail blood and purification of mature P. berghei schizonts sufficient for a single transfection experiment are described.162Research highlights► Small-scale overnight culture and purification of schizonts of P. berghei are performed in 1.5-mL microcentrifuge tubes. ► The purified schizonts can be used in successful transfection experiments. ► This small-scale protocol provides significant advantages, namely reduction of parasite sample, laboratory consumables and mice for transfection experiments.