Trypanosoma brucei gambiense: HMI-9 medium containing methylcellulose and human serum supports the continuous axenic in vitro propagation of the bloodstream form

• Published in: Experimental parasitology 2011-07, Vol.128 (3), p.285-290
• Main Authors: Van Reet, N., Pyana, P.P., Deborggraeve, S., Büscher, P., Claes, F.
• Format: Article
• Language: English
• Subjects: African trypanosomiasis; Animals; axenic culture; Biological and medical sciences; blood flow; Bloodstream form; Culture Media - chemistry; Democratic Republic of the Congo; Female; fetal bovine serum; Freezing; Fundamental and applied biological sciences. Psychology; HMI-9; Human protozoal diseases; Human serum; Humans; In vitro; Infectious diseases; Life cycle. Host-agent relationship. Pathogenesis; Medical sciences; Methylcellulose; Mice; Parasitic diseases; parasitology; Protozoa; Protozoal diseases; Serum; Trypanosoma brucei; Trypanosoma brucei gambiense; Trypanosoma brucei gambiense - classification; Trypanosoma brucei gambiense - growth & development; Trypanosoma brucei gambiense - physiology; Trypanosoma gambiense; Trypanosomiasis; Trypanosomiasis, African - parasitology
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2011.02.018

[Display omitted] ► HMI-9 medium with methylcellulose is used for culture of pleomorphic T.b. brucei. ► We investigated the suitability of this medium for bloodstream form T.b. gambiense. ► Additional supplementation of methylcellulose-HMI-9 with human serum is critical. Trypanosoma brucei (T.b.) gambiense causes the chronic form of human African trypanosomiasis or sleeping sickness. One of the major problems with studying T.b. gambiense is the difficulty to isolate it from its original host and the difficult adaptation to in vivo and in vitro mass propagation. The objective of this study was to evaluate if an established method for axenic culture of pleomorphic bloodstream form T.b. brucei strains, based on methylcellulose containing HMI-9 medium, also facilitated the continuous in vitro propagation of other bloodstream form Trypanozoon strains, in particular of T.b. gambiense. Bloodstream form trypanosomes from one T.b. brucei, two T.b. rhodesiense, one T. evansi and seven T.b. gambiense strains were isolated from mouse blood and each was concurrently cultivated in liquid and methylcellulose-containing HMI-9 based medium, either with or without additional human serum supplementation, for over 10 consecutive sub passages. Although HMI-9 based medium supplemented with 1.1% (w/v) methylcellulose supported the continuous cultivation of all non-gambiense strains better than liquid media could, the in vitro cultivation of all gambiense strains was only achieved in HMI-9 based medium containing 1.1% (w/v) methylcellulose, 15% (v/v) fetal calf serum and 5% (v/v) heat-inactivated human serum.