Short communication: cultivation of bloodstream forms of Trypanosoma brucei and T. evansi in a serum-free medium.
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 supplemental 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 alpha 2-macroglobulin, bovine beta-lipoprotein, d-biotin, retinol, beta-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 x 10(6) cells/ml; 10.6 hours and 2.2 x 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.
Hirumi
H
H
Research Center for Protozoan Molecular Immunology, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
Martin
S
S
Hirumi
K
K
Inoue
N
N
Kanbara
H
H
Saito
A
A
Suzuki
N
N
D000818:Animals; D002417:Cattle; D016895:Culture Media, Serum-Free / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D018512:Parasitemia / Q000469:parasitology; D010274:Parasitology / Q000379:methods; D012710:Serum Albumin, Bovine; D014345:Trypanosoma / Q000254:growth & development* / Q000302:isolation & purification; D014346:Trypanosoma brucei brucei / Q000254:growth & development* / Q000302:isolation & purification; D014352:Trypanosomiasis / Q000469:parasitology; D014353:Trypanosomiasis, African / Q000469:parasitology
D016895:Culture Media, Serum-Free; D012710:Serum Albumin, Bovine