Synchronization of Trypanosoma brucei by Counter-Flow Centrifugal Elutriation

Benz, Corinna and Urbaniak, Michael D (2022) Synchronization of Trypanosoma brucei by Counter-Flow Centrifugal Elutriation. In: Cell-Cycle Synchronization. Methods in Molecular Biology . Humana Press, New York, pp. 137-144. ISBN 9781071627358

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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.

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20 Sep 2022 09:05
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20 Sep 2022 13:50