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  • 标题:A simple 3D cellular chemotaxis assay and analysis workflow suitable for a wide range of migrating cells
  • 本地全文:下载
  • 作者:Sai P. Visweshwaran ; Tanja Maritzen
  • 期刊名称:MethodsX
  • 印刷版ISSN:2215-0161
  • 电子版ISSN:2215-0161
  • 出版年度:2019
  • 卷号:6
  • 页码:2807-2821
  • DOI:10.1016/j.mex.2019.11.001
  • 语种:English
  • 出版社:Elsevier
  • 摘要:Graphical abstractDisplay OmittedAbstractCellular migration plays a crucial role within multicellular organisms enabling organ development, wound healing and efficient immune responses, but also metastasis. Therefore, it is crucial to dissect the underlying mechanisms. Directed migration and invasion are most efficient in response to chemotactic signals. To study cell migration and chemotactic responses, current experimental setups use either simplified 2D, tissue-mimetic 3D (e.g. collagen matrices) orin vivoenvironments. While thein vivoexperiments are closest to the real physiological situation, they require animal experiments and are thus ill-suited for screening purposes. 3D matrices, on the other hand, can mimicin vivoconditions in many respects thus serving as instructive settings for the initial dissection of cell migration and chemotaxis. However, performing 3D chemotaxis assays has its limitations due to the delicate nature of most available setups and the tedious and time-consuming manual quantification process. Here, we present•A method for the easy construction of a chemotaxis chamber suitable for the analysis of large cell numbers.•A procedure to quantify their migration automatically with minimal input required by the experimenter.•Both successfully validated by analyzing the 3D chemotaxis of highly migratory primary dendritic cells and the invasive MDA-MB-231 cancer cells.
  • 关键词:Migration;Motility;Collagen;Dendritic cells;Cancer;Directionality;Automated cell tracking;Cell speed;Mean Squared Displacement (MSD);Three dimensional cell migration;TrackMate;T cells
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