High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis.
High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis.
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DOI：
10.1038/srep27301
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发表时间：
2016-06-13
影响因子：
4.6
通讯作者：
Yoon E
中科院分区：
文献类型：
--
作者：
Chen YC;Ingram PN;Fouladdel S;McDermott SP;Azizi E;Wicha MS;Yoon E
相当多的证据表明，许多恶性肿瘤是由显示干细胞特性的细胞区室驱动的。肿瘤干细胞样细胞（CSCs）可以通过细胞表面标志物的表达或酶活性来鉴定，但这些方法受到CSCs表型异质性和可塑性的限制。已经开发了基于体外球体形成的替代表型方法，但它通常是劳动密集型和低通量的。在这项工作中，我们提出了一个1,024微室微流控平台的单细胞衍生球的形成。利用流体动力学捕获方案，超过70%的微室仅捕获一个细胞，允许监测来自异质癌细胞群体的球体形成以鉴定CSC。可以使用定制的96-基因组取回并解离单细胞衍生的球体以用于单细胞分析，以探测克隆CSC球体内的异质性。该微流体平台为CSC鉴定和下游克隆分析提供了可靠和高通量的球体形成。
Considerable evidence suggests that many malignancies are driven by a cellular compartment that displays stem cell properties. Cancer stem-like cells (CSCs) can be identified by expression of cell surface markers or enzymatic activity, but these methods are limited by phenotypic heterogeneity and plasticity of CSCs. An alternative phenotypic methodology based on in-vitro sphere formation has been developed, but it is typically labor-intensive and low-throughput. In this work, we present a 1,024-microchamber microfluidic platform for single-cell derived sphere formation. Utilizing a hydrodynamic capturing scheme, more than 70% of the microchambers capture only one cell, allowing for monitoring of sphere formation from heterogeneous cancer cell populations for identification of CSCs. Single-cell derived spheres can be retrieved and dissociated for single-cell analysis using a custom 96-gene panel to probe heterogeneity within the clonal CSC spheres. This microfluidic platform provides reliable and high-throughput sphere formation for CSC identification and downstream clonal analysis.