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Synchronization of Trypanosoma brucei by Counter-Flow Centrifugal Elutriation

Research output: Contribution in Book/Report/Proceedings - With ISBN/ISSNChapter (peer-reviewed)peer-review

Published
Publication date1/09/2022
Host publicationCell-Cycle Synchronization: Methods and Protocols
EditorsZhixiang Wang
Place of PublicationNew York
PublisherHumana Press
Pages137-144
Number of pages8
ISBN (Electronic)9781071627365
ISBN (Print)9781071627358
<mark>Original language</mark>English

Publication series

NameMethods in Molecular Biology
Volume2579
ISSN (Print)1940-6029

Abstract

Centrifugal counter-flow elutriation is a non-invasive technique that separates cells based on their hydrodynamic volume in a specialized centrifugation chamber that allows the application of a counter-flow of buffer to oppose sedimentation. Here, we report a centrifugal counter-flow elutriation protocol for Trypanosoma brucei cells that is able to rapidly isolate highly enriched G1 subpopulations (>95%) of synchronized cells. The cells obtained are viable and proliferate without lag, allowing subsequent cell cycle phases to be obtained by continued culture. The synchronized cell cultures obtained by this process have uniform DNA content, a narrow size distribution, undergo synchronous division, and maintain synchrony into subsequent cell cycles.