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PMID: 2616571 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Long-term culture and cloning system for Trypanosoma brucei gambiense bloodstream forms in semi-defined medium in vitro.

Parasitology research ·Vol. 76 ·No. 2 ·1989-00-00 ·Pages 93-7

Yabu Y, Takayanagi T, Sato S

Abstract

A new semi-defined medium extremely useful for the long-term cultivation and cloning of Trypanosoma b. gambiense (Wellcome strain) bloodstream forms is described. Bloodstream forms could be continuously grown in 25 mM HEPES-buffered D-MEM supplemented with 10 microM bathocuproine sulfonate (BCS), 100 microM cysteine, and 20% heat-inactivated fetal calf serum at 37 degrees C in vitro. Under these culture conditions, T. b. gambiense bloodstream forms increased in number up to 2-3 x 10(6) trypanosomes/ml by day 3 after initiation of the culture. The trypanosomes maintained in this culture system for 200 days retained their infectivity for mice. Morphologically, they were long and slender, and a surface coat was evident on the cell surface and flagellar membrane. In vitro cloning with single bloodstream forms of T. b. gambiense could be achieved with high efficiency.

MeSH Terms
Animals Culture Media Cysteine/metabolism Indicators and Reagents Mice Microscopy, Electron Oxidation-Reduction Phenanthrolines/metabolism Trypanosoma brucei gambiense/growth & development,pathogenicity,ultrastructure Virulence
Chemicals
Culture Media Indicators and Reagents Phenanthrolines bathocuproine sulfonate Cysteine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Yabu Y
Department of Medical Zoology, Nagoya City University, Medical School, Japan.
Takayanagi T
Sato S
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Article Info
Journal
Parasitology research
Abbr.
Parasitol Res
ISSN
0932-0113
DOI
10.1007/BF00930828
Published
1989-00-00
Pages
93-7
Language
English
Region
Germany
NLM ID
8703571
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00930828 View journal details
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