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Trypanosoma brucei : Differentiation of in Vitro‐Grown Bloodstream Trypomastigotes into Procyclic Forms 1
Author(s) -
SIMPSON AGDA M.,
HUGHES DALLAS,
SIMPSON LARRY
Publication year - 1985
Publication title -
the journal of protozoology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 0022-3921
DOI - 10.1111/j.1550-7408.1985.tb03100.x
Subject(s) - in vitro , trypanosoma brucei , microbiology and biotechnology , cell culture , biology , cell division , cellular differentiation , cell , biochemistry , genetics , gene
Trypanosoma brucei strain 366D trypomastigotes grown at 37°C in the presence of a human fibroblast cell line formed foci underneath the feeder cells whereas trypanosomes grown in the presence of a human epithelial cell line grew only in the culture supernatant. A culture system was developed to study the differentiation of bloodstream trypomastigotes grown in the epithelial cell system into procyclic trypomastigotes at 27°C. The morphological differentiation into the procyclic form was complete by 48 h. Cell division did not occur until 30–40 h after transfer to 27°C. Various characteristics of this system were examined, including the effect of the feeder layer, the type of medium, the presence of the metabolites cis‐aconitate and citrate, the preadaptation period, and the trypanosome cell concentration. The respiration of the recently differentiated procyclic cells was less sensitive to inhibition by CN‐than that of established procyclic forms, implying a delayed appearance of complete mitochondrial oxidative pathways. This trypanosome differentiation system has the advantage that the animal host is not needed and the entire process is carried out in in vitro culture.