Single-Cell-Derived Tumor-Sphere Formation and Drug-Resistance Assay Using an Integrated Microfluidics
Single-Cell-Derived Tumor-Sphere Formation and Drug-Resistance Assay Using an Integrated Microfluidics
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使用集成微流体进行单细胞衍生肿瘤球形成和耐药性测定
DOI：
10.1021/acs.analchem.9b01084
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发表时间：
2019-07-02
影响因子：
7.4
通讯作者：
Fan, Shih-Kang
中科院分区：
文献类型：
--
作者：
Pang, Long;Ding, Jing;Fan, Shih-Kang
大量证据表明，肿瘤干细胞(CSCs)在促进肿瘤进展、转移和耐药方面起着重要作用。在不损害细胞生物学特性的情况下，单细胞来源的肿瘤球体是CSCs鉴定和研究的鼓舞人心的选择。尽管已经开发了几种基于单细胞的微流控方法用于CSCs的研究，但澄清细胞的生物力学(如大小和变形性)与干细胞(如肿瘤球体的形成和耐药性)之间的联系仍然具有挑战性。在这里，我们提出了一个集成的微流体平台，用于分析单细胞来源的肿瘤球体的形成和耐药性。使用该装置可以有效地形成来自不同生物力学(大小和/或变形)的单细胞的肿瘤球体。为了验证微流控平台的能力，通过评估具有不同生物力学特性的单个胶质母细胞瘤细胞的单个细胞衍生球体的形成，进行了概念验证实验。此外，通过与长春新碱共培养来确定研究这些单细胞衍生微球的化疗疗程。结果表明，肿瘤细胞的生物力学与单细胞衍生球体的形成有关，即较小和/或更易变形的肿瘤细胞由单细胞衍生球体的形成定义为更像干细胞，而不是更突出和/或更不变形的肿瘤细胞。此外，由单个较小和/或较易变形的肿瘤细胞衍生的肿瘤球体比更突出和/或较不易变形的肿瘤细胞具有更高的耐药性。我们的装置根据肿瘤细胞不同的生物力学特性，为单细胞衍生球的形成提供了一种新的方法。此外，它还为CSC的识别和单细胞水平的下游分析提供了一种新的方法。
Considerable evidence points to cancer stem-like cells (CSCs) as responsible for promoting progression, metastasis, and drug resistance. Without damage to the cell biological properties, single-cell-derived tumor-sphere is encouraging options for CSCs identification and studies. Although several single cell-based microfluidic methods have been developed for CSCs studies, clarifying liaison between the biomechanics of cells (such as size and deformability) and stem (such as tumor-sphere formation and drug resistance) remains challenging. Herein, we present a platform of integrated microfluidics for the analysis of single-cell-derived tumorsphere formation and drug resistance. Tumor-spheres derived from different biomechanics (size and/or deformation) single-cells could be formed efficiently using this device. To demonstrate the microfluidic-platform capability, a proof-of-concept experiment was implemented by evaluating single-cell-derived sphere formation of single glioblastoma cells with different biomechanics. Additionally, a course of chemotherapy to study these single- cell-derived spheres was determined by coculture with vincristine. The results indicate that tumor cell biomechanics is associated with single-cell-derived spheres formation; that is, smaller and/or more deformable tumor cells are more stem-like defined by the formation of single-cell-derived spheres than more prominent and/or lesser deformable tumor cells. Also, tumor- spheres derived from single small and/or more deformable tumor cell have higher drug resistance than more prominent and/or less deformable tumor cells. Our device offers a new approach for single-cell-derived sphere formation according to tumor cell different biomechanical properties. Furthermore, it offers a new method for CSC identification and downstream analysis on a single-cell level.