Screening Therapeutic Agents Specific to Breast Cancer Stem Cells Using a Microfluidic Single-Cell Clone-Forming Inhibition Assay
Screening Therapeutic Agents Specific to Breast Cancer Stem Cells Using a Microfluidic Single-Cell Clone-Forming Inhibition Assay
复制标题
使用微流体单细胞克隆形成抑制试验筛选乳腺癌干细胞特异性治疗剂。
DOI：
10.1002/smll.201901001
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发表时间：
2020-03-01
期刊：
影响因子：
13.3
通讯作者：
Liu, Dayu
中科院分区：
文献类型：
--
作者：
Lin, Dongguo;Li, Peiwen;Liu, Dayu
肿瘤干细胞(CSCs)特异性试剂的筛选给传统的细胞分析带来了巨大的挑战，因为很难准备好准备用于药物测试的CSCs。为了克服这一局限性，开发了一种用于筛选乳腺癌干细胞特异性药物的微流控单细胞实验。这种检测方法利用了CSC的单细胞克隆形成能力，这种能力可以被CSC靶向药物特异性地抑制。单细胞分析在具有3840个细胞捕获单元的阵列的微流控芯片上进行；通过将细胞悬浮液流入微芯片很容易形成单细胞阵列。实现了接近60%的单细胞捕获率，从而允许在一次测试中分析2000多个单细胞。在长期悬浮培养中，只有一小部分细胞存活并形成肿瘤球体。MCF-7、MDA-MB-231和T47D细胞的克隆形成率分别为1.67%、5.78%和5.24%。克隆形成抑制试验通过将单细胞阵列暴露于一组抗癌剂来进行。CSC靶向药物对单细胞克隆形成有完全抑制作用，而非靶向药物则表现出不完全抑制作用。由此产生的微流控单细胞分析具有高效筛选CSC特异性药物的潜力，为个体化肿瘤治疗提供了新的工具。
Screens of cancer stem cells (CSCs)-specific agents present significant challenges to conventional cell assays due to the difficulty in preparing CSCs ready for drug testing. To overcome this limitation, developed is a microfluidic single-cell assay for screening breast cancer stem cell-specific agents. This assay takes advantage of the single-cell clone-forming capability of CSCs, which can be specifically inhibited by CSC-targeting agents. The single-cell assay is performed on a microfluidic chip with an array of 3840 cell-capturing units; the single-cell arrays are easily formed by flowing a cell suspension into the microchip. Achieved is a single cell-capture rate of approximate to 60% thus allowing more than 2000 single cells to be analyzed in a single test. Over long-term suspension culture, only a minority of cells survive and form tumorspheres. The clone-formation rate of MCF-7, MDA-MB-231, and T47D cells is 1.67%, 5.78%, and 5.24%, respectively. The clone-forming inhibition assay is conducted by exposing the single-cell arrays to a set of anticancer agents. The CSC-targeting agents show complete inhibition of single-cell clone formation while the nontargeting ones show incomplete inhibition effects. The resulting microfluidic single-cell assay with the potential to screen CSC-specific agents with high efficiency provides new tools for individualized tumor therapy.