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PMID: 1528898 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

High-efficiency clonal growth of bloodstream- and insect-form Trypanosoma brucei on agarose plates.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 89 ·No. 18 ·1992-09-15 ·Pages 8818-21

Carruthers VB, Cross GA

Abstract

This report describes a method for growing both bloodstream- and procyclic-form Trypanosoma brucei as colonies on agarose plates. Procyclic colonies, which took 2 weeks to develop, grew with approximately 17% plating efficiency on SDM-79/0.65% agarose supplemented with 20% (vol/vol) conditioned medium. Bloodstream forms were adapted to in vitro growth in liquid HMI-9 medium and then spread on HMI-9/0.65% agarose plates, where they grew to visible colonies in 3-5 days. Plating efficiencies were from 3 to 80%, depending upon the trypanosome variant and experiment. Colonies were proven to be the result of growth from a single cell and contained approximately 10(6) cells at maturity. Colonies were transferred to filters and probed for multicopy and single-copy genes. Potential uses of this method in conjunction with classical and reverse genetic approaches to studying trypanosomes are discussed.

MeSH Terms
Animals Cell Division Culture Media In Vitro Techniques Sepharose Trypanosoma brucei brucei/growth & development
Chemicals
Culture Media Sepharose
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Carruthers V B
Laboratory of Molecular Parasitology, Rockefeller University, New York, NY 10021.
Cross G A
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31 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.89.18.8818
Published
1992-09-15
Pages
8818-21
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC50012
Subset
IM
Grants
NIAID NIH HHS · AI21729 · United States
External Links
PubMed source DOI: 10.1073/pnas.89.18.8818 PMC Full Text View journal details
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