Trypsin-stimulated transformation of Trypanosoma brucei gambiense bloodstream forms to procyclic forms in vitro

The effect of trypsin treatment on the transformation of monomorphic Trypanosoma b. gambiense (Wellcome strain) bloodstream forms to procyclic forms was studied in HEPES-buffered RPMI 1640 medium supplemented with 20% inactivated fetal calf serum in the presence of GA-1 cells as feeder layers at 27 degrees C. In this system, 35%-40% of the bloodstream forms transformed to procyclic forms within 24 h, and over 95% of the trypanosomes changed into procyclic forms by day 3 after initiation of the culture. Established cultures of procyclic forms yielded up to 1.5-2 x 10(7) trypanosomes/ml. However, transformation of nontreated and inhibited trypsin-treated bloodstream forms were prolonged compared to trypsin-treated populations. In this experiment, the first procyclic forms could be detected on day 7 after initiation of the culture and transformation was complete within 15 days. The transformation of T. b. gambiense from bloodstream to procyclic forms required the living GA-1 cells as feeder layer cells, but established cultures of procyclic forms could be maintained in the culture medium without feeder cells for more than 300 days.