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A quick, simple method for purifying Leishmania mexicana amastigotes in large numbers

Published online by Cambridge University Press:  06 April 2009

D. T. Hart ,
K. Vickerman  and
G. H. Coombs
D. T. Hart
Affiliation:
Department of Zoology, University of Glasgow, Glasgow G12 8QQ
K. Vickerman
Affiliation:
Department of Zoology, University of Glasgow, Glasgow G12 8QQ
G. H. Coombs
Affiliation:
Department of Zoology, University of Glasgow, Glasgow G12 8QQ
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Summary

A rapid method for the bulk isolation of purified Leishmania mexicana mexicana amastigotes from parasite-induced lesions in experimentally infected mice is described. The procedure includes purification steps based on differences in net cell charge, lysis susceptibility and buoyant density between parasite and host cells. Yields of up to 2 × 1010 untransformed amastigotes with minimal contamination with host cells and cell debris can be obtained. At least 90 % of the purified amastigotes are viable as judged by light and electron microscopy, the staining of their lysosomes with acridine orange, their ability to transform to promastigotes and their infectivity to macrophages in vivo and in vitro.

Type
Research Article
Information
Parasitology , Volume 82 , Issue 3 , June 1981 , pp. 345 - 355
Copyright
Copyright © Cambridge University Press 1981

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