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The Axenic Cultivation of Insect Forms of Trypanosoma (Duttonella) vivax and Development to the Infective Metacyclic Stage
Author(s) -
GUMM IAN D.
Publication year - 1991
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.1550-7408.1991.tb04424.x
Subject(s) - axenic , trypanosoma vivax , biology , axenic culture , laboratory flask , proline , microbiology and biotechnology , glutamine , trypanosoma , biochemistry , chemistry , bacteria , virology , amino acid , genetics
A method for axenic cultivation of epimastigote and metacyclic forms of Trypanosoma (Duttonella) vivax at 27°C in vitro is described. Iscove's medium was supplemented with specific concentrations of foetal bovine serum, L‐proline, L‐glutamine, hypoxanthine, adenosine, pyruvate, and 2‐mercaptoethanol. Bloodstream form parasites rapidly transformed into epimastigote forms that grew as surface‐adherent colonies in plastic culture flasks. Transformation of epimastigotes to metacyclic forms was first observed 9–12 days after initiation of cultures. Percentages of metacyclics varied: East African T. vivax ranged up to 40% and West African T. vivax ranged up to 24%. Subcultures were made at two‐week intervals and maintained for several months. Transformation of bloodstream forms to epimastigotes depended on initial attachment to the bottom of culture flasks and the presence of L‐proline. The number and maturity of metacyclic forms was influenced by the concentrations of foetal bovine serum, L‐proline, L‐glutamine, and 2‐mercaptoethanol. Trypanosomes from cultures were cryopreserved, revived, and used to re‐establish fresh axenic cultures. These results represent a significant advance in cultivation of T. vivax insect forms that should enable studies to be accomplished on metabolism, differentiation, and pharmacology of this parasitic protozoan, free from the influence of extraneous cells.