New synchronization method for Plasmodium falciparum

Malar J. 2010 Jun 17:9:170. doi: 10.1186/1475-2875-9-170.

Abstract

Background: Plasmodium falciparum is usually asynchronous during in vitro culture. Although various synchronization methods are available, they are not able to narrow the range of ages of parasites. A newly developed method is described that allows synchronization of parasites to produce cultures with an age range as low as 30 minutes.

Methods: Trophozoites and schizonts are enriched using Plasmion. The enriched late stage parasites are immobilized as a monolayer onto plastic Petri dishes using concanavalin A. Uninfected erythrocytes are placed onto the monolayer for a limited time period, during which time schizonts on the monolayer rupture and the released merozoites invade the fresh erythrocytes. The overlay is then taken off into a culture flask, resulting in a highly synchronized population of parasites.

Results: Plasmion treatment results in a 10- to 13-fold enrichment of late stage parasites. The monolayer method results in highly synchronized cultures of parasites where invasion has occurred within a very limited time window, which can be as low as 30 minutes. The method is simple, requiring no specialized equipment and relatively cheap reagents.

Conclusions: The new method for parasite synchronization results in highly synchronized populations of parasites, which will be useful for studies of the parasite asexual cell cycle.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Concanavalin A
  • Culture Media
  • Culture Techniques / methods*
  • Erythrocytes / parasitology*
  • Gelatin
  • Mitogens
  • Plasma Substitutes
  • Plasmodium falciparum / growth & development*
  • Plasmodium falciparum / isolation & purification
  • Schizonts / parasitology
  • Trophozoites / parasitology

Substances

  • Culture Media
  • Mitogens
  • Plasma Substitutes
  • Concanavalin A
  • plasmion
  • Gelatin