Division of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, Lancaster, UK.
Methods Mol Biol. 2022;2579:137-144. doi: 10.1007/978-1-0716-2736-5_11.
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.
离心逆流淘洗是一种非侵入性技术，它根据细胞的流体力学体积在专门的离心室中分离细胞，该离心室允许应用缓冲液的逆流来对抗沉降。在这里，我们报告了一种用于布氏锥虫细胞的离心逆流淘洗方案，该方案能够快速分离高度富集的 G1 亚群（>95%）的同步细胞。获得的细胞是有活力的，并且没有延迟地增殖，从而允许通过继续培养获得后续的细胞周期阶段。通过该过程获得的同步细胞培养物具有均匀的 DNA 含量、狭窄的大小分布、进行同步分裂，并在随后的细胞周期中保持同步。
