Ex Vivo Maturation Assay for Testing Antimalarial Sensitivity of Rodent Malaria Parasites.
Ex vivo assay systems provide a powerful approach to studying human malaria parasite biology and to testing antimalarials. For rodent malaria parasites, short-term in vitro culture and ex vivo antimalarial susceptibility assays are relatively cumbersome, relying on in vivo passage for synchronization, since ring-stage parasites are an essential starting material. Here, we describe a new approach based on the enrichment of ring-stage Plasmodium berghei, P. yoelii, and P. vinckei vinckei using a single-step Percoll gradient. Importantly, we demonstrate that the enriched ring-stage parasites develop synchronously regardless of the parasite strain or species used. Using a flow cytometry assay with Hoechst and ethidium or MitoTracker dye, we show that parasite development is easily and rapidly monitored. Finally, we demonstrate that this approach can be used to screen antimalarial drugs.
Singapore Immunology Network (SIgN), A*STAR, Biopolis, Singapore.|Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System, Singapore.
Zi Wei
ZW
Chang
Singapore Immunology Network (SIgN), A*STAR, Biopolis, Singapore.|Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System, Singapore.
Benoit
B
Malleret
Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System, Singapore.|Department of Microbiology and Immunology, Otago University, Dunedin, New Zealand.
Bruce
B
Russell
Singapore Immunology Network (SIgN), A*STAR, Biopolis, Singapore renia_laurent@immunol.a-star.edu.sg carla_claser@immunol.a-star.edu.sg.|Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System, Singapore.
Laurent
L
Rénia
Singapore Immunology Network (SIgN), A*STAR, Biopolis, Singapore renia_laurent@immunol.a-star.edu.sg carla_claser@immunol.a-star.edu.sg.
Carla
C
Claser
D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D008288:Malaria / Q000188:drug therapy / Q000469:parasitology*; D008297:Male; D008810:Mice, Inbred C57BL; D008826:Microbial Sensitivity Tests; D010961:Plasmodium / Q000187:drug effects* / Q000472:pathogenicity / Q000502:physiology*; D010962:Plasmodium berghei / Q000187:drug effects / Q000472:pathogenicity / Q000502:physiology
D000962:Antimalarials