Cultivation of the life cycle stages of Trypanosoma brucei sspp

A culture system was devised for the production of the various stages in the developmental cycle of Trypanosoma brucei brucei and T. b. rhodesiense. The bloodstream forms were grown at 37 degrees C on a feeder layer of fibroblasts from embryos of Microtus montanus or CD-1 mice in HEPES-buffered Minimum Essential Medium with Earle's salts, supplemented with 15% heat-inactivated rabbit serum. When they were transferred to HEPES-buffered Cunningham's medium and incubated at 27 degrees C, they transformed into procyclic trypomastigotes, some of which developed into epimastigote and metacyclic forms in the presence of explants of Glossina morsitans or Phormia regina. Stages infective to mice were produced in cultures of procyclic forms grown with Anopheles gambiae cells in medium consisting of a mixture of 3 volumes of Cunningham's medium and 1 volume of Anopheles cell medium. In vitro-produced metacyclic trypanosomes were used to initiate cultures of bloodstream forms.