Synchronization of Trypanosoma brucei by Counter-Flow Centrifugal Elutriation.
Synchronization of Trypanosoma brucei by Counter-Flow Centrifugal Elutriation.
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通过逆流离心淘洗同步布氏锥虫。
DOI：
10.1007/978-1-0716-2736-5_11
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Benz C
中科院分区：
文献类型：
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作者：
Benz C
离心逆流洗脱是一种非侵入性技术，它根据细胞的流体动力学体积在专门的离心室中分离细胞，允许应用逆流缓冲液来防止沉淀。在这里，我们报告了一种离心逆流洗脱方案，该方案能够快速分离高度富集的G1亚群（bb0 95%）的同步细胞。获得的细胞是有活力的，无延迟增殖，允许通过继续培养获得后续的细胞周期阶段。通过该方法获得的同步细胞培养物DNA含量均匀，大小分布窄，分裂同步，并在随后的细胞周期中保持同步。
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 forTrypanosoma bruceicells 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.