Fluorescence-Activated Cell Sorting Analysis of Heterotypic Cell-in-Cell Structures.
Fluorescence-Activated Cell Sorting Analysis of Heterotypic Cell-in-Cell Structures.
复制标题
异型细胞内结构的荧光激活细胞分选分析
DOI：
10.1038/srep09588
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发表时间：
2015-04-27
影响因子：
4.6
通讯作者：
Sun Q
中科院分区：
文献类型：
--
作者：
He M;Huang H;Wang M;Chen A;Ning X;Yu K;Li Q;Li W;Ma L;Chen Z;Wang X;Sun Q
细胞内结构(cell-in-cell Structures，CICs)是指细胞内部存在一个或多个活性细胞，近年来被发现在发育、免疫稳态和肿瘤发生等方面发挥着重要作用。与人们对这一独特现象的日益关注不相适应，目前还缺乏可靠的高通量定量和系统研究方法。在这里，我们报告了一种基于流式细胞术的方法，用于快速分析和分选在淋巴细胞和肿瘤细胞之间形成的异型CICs。该方法用荧光染料标记细胞，用流式细胞仪进行荧光激活细胞分选，优化了减少细胞倍体的条件，可获得高纯度(~gt；95%)的CICs。利用这种方法，我们分析了不同细胞对之间CICs的形成，发现内化效应细胞和吞噬靶细胞的因素都会影响异型CICs的形成。因此，基于流式细胞术的FACS分析将成为推动CICs系统研究的高通量方法。
Cell-in-cell structures (CICs), characterized by the presence of one or more viable cells inside another one, were recently found important player in development, immune homeostasis and tumorigenesis etc. Incompatible with ever-increasing interests on this unique phenomenon, reliable methods available for high throughput quantification and systemic investigation are lacking. Here, we report a flow cytometry-based method for rapid analysis and sorting of heterotypic CICs formed between lymphocytes and tumor cells. In this method, cells were labeled with fluorescent dyes for fluorescence-activated cell sorting (FACS) by flow cytometry, conditions for reducing cell doublets were optimized such that high purity (>95%) of CICs could be achieved. By taking advantage of this method, we analyzed CICs formation between different cell pairs and found that factors from both internalized effector cells and engulfing target cells affect heterotypic CICs formation. Thus, flow cytometry-based FACS analysis would serve as a high throughput method to promote systemic researches on CICs.