Premium
Cultivation of bloodstream forms of Trypanosoma brucei and T. evansi in a serum‐free medium
Author(s) -
Hirumi H.,
Martin S.,
Hirumi K.,
Inoue N.,
Kanbara H.,
Saito A.,
Suzuki N.
Publication year - 1997
Publication title -
tropical medicine and international health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.056
H-Index - 114
eISSN - 1365-3156
pISSN - 1360-2276
DOI - 10.1046/j.1365-3156.1997.d01-268.x
Subject(s) - biology , fetal bovine serum , bovine serum albumin , hypoxanthine , trypanosoma evansi , population , biochemistry , microbiology and biotechnology , trypanosomiasis , virology , in vitro , medicine , environmental health , enzyme
Bloodstream forms of Trypanosoma brucei and T. evansi have been cultivated in an axenic culture system by using Iscove's modified DMEM‐based HMI‐9 medium supplemented with bathocuproinedisulphonic acid, l ‐cysteine, hypoxanthine, 2‐mercaptoethanol, pyruvate, thymidine and 10% fetal bovine or adult horse serum. We developed a serum‐free medium (HMI‐244) in which serum in HMI‐9 was replaced by fatty acid‐free bovine serum albumin, bovine α 2 ‐macroglobulin, bovine β‐lipoprotein, d‐biotin, retinol, β‐alanine, l‐anserine nitrate salt, l‐ornithine hydrochloride, O‐phosphorylethanolamine, sarcosine, taurine, adenosine‐ 5′‐triphosphate, 2′‐deoxycytidine‐5′‐monophosphate, 2′‐deoxyguanosine‐5′‐monophosphate, and 5‐methyltetrahydrofolic acid. Maximum cell densities and population doubling times were 2.6×10 6 cells/ml; 10.6 hours and 2.2×10 6 cells/ml; 10.9 hours for T. brucei and T. evansi , respectively. Bloodstream forms continued to proliferate in the serum‐free cultures for more than 90 days and the trypomastigotes retained their morphological characteristics and infectivity to mice. If validated, this serum‐free medium may help reduce future interlaboratory variability in biochemical, immunological, molecular biological and drug sensitivity studies on these parasites.