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

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 p...

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Bibliographic Details
Published inParasitology Vol. 82; no. Pt 3; p. 345
Main Authors Hart, D T, Vickerman, K, Coombs, G H
Format Journal Article
LanguageEnglish
Published England 01.06.1981
Subjects
Animals
Centrifugation, Isopycnic
Chromatography, Ion Exchange
Female
Leishmania - growth & development
Leishmania - isolation & purification
Leishmania - physiology
Leishmaniasis - parasitology
Mice
Parasitology - methods
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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 x 10(10) 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.
ISSN:0031-1820
DOI:10.1017/S0031182000066889