Hydroxyurea-induced synchronisation of bloodstream stage Trypanosoma brucei.

Synchronisation of the Trypanosoma brucei cell cycle proved elusive for many years. A recent report demonstrated that synchronisation of procyclic form cells was possible following treatment with hydroxyurea. Here, that work is extended to the disease-relevant, mammalian-infective bloodstream stage trypanosome. Treatment of bloodstream stage Lister 427 T. brucei cells growing in vitro with 10 microg ml(-1) hydroxyurea for 6h led to an enrichment of cells in S phase. Following removal of the drug, cells proceeded uniformly through one round of the cell cycle, providing a much needed tool to enrich for specific cell cycle stages, in a manner similar to hydroxyurea treatment of procyclic form T. brucei.