Microfluidics 3D gel-island chip for single cell isolation and lineage-dependent drug responses study.
Microfluidics 3D gel-island chip for single cell isolation and lineage-dependent drug responses study.
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DOI：
10.1039/c6lc00081a
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发表时间：
2016-07-07
期刊：
影响因子：
6.1
通讯作者：
Yoon E
中科院分区：
文献类型：
--
作者：
Zhang Z;Chen YC;Cheng YH;Luan Y;Yoon E
细胞外基质（ECM）中的3D细胞培养不仅为细胞成分提供结构支持，而且还为组织中的各种重要细胞功能启动调节生化线索，在理解癌症病理学和药物测试中变得越来越重要。尽管ECM凝胶已用于批量和芯片上的细胞培养，但以前的研究集中在集体细胞行为而不是单细胞异质性。为了跟踪每个细胞的行为，我们开发了一种凝胶岛芯片，它可以形成数千个包含被所需ECM包裹的单细胞的岛。通过泊松分布优化，该装置可以达到每个岛的确切单个细胞数的34%的捕获效率。在凝胶岛中可以实现良好的培养基交换率和高细胞活力。岛中的细胞可以自动计数用于高通量分析。作为概念证明，我们监测了单个Notch+（干细胞样）T47 D乳腺癌细胞的增殖和分化。发现3D胶原凝胶环境通过更好的自我更新和去分化（Notch−到Notch+转变）有利于干细胞样表型。更有趣的是，我们发现Notch-去分化的细胞比Notch+细胞对阿霉素和顺铂更耐药。结合三维ECM培养和单细胞分辨率，所提出的平台可以使用少量的药物和试剂自动分析数百个细胞的单个细胞行为。
3D cell culture in the extracellular matrix (ECM), which not only provides structural support to cellular constituents, but also initiates regulatory biochemical cues for a variety of important cell functions in tissue, has become more and more important in understanding cancer pathology and drug testing. Although the ECM-gel has been used in cell culture both in bulk and on-chip, previous studies focused on collective cell behavior rather than single-cell heterogeneity. To track the behavior of each individual cell, we have developed a gel-island chip, which can form thousands of islands containing single cells encapsulated by the desired ECM. Optimized by Poisson's distribution, this device can attain 34% capture efficiency of the exact number of single cells per island. A good culture media exchange rate and high cell viability can be achieved in the gel-islands. The cells in the islands can be automatically counted for high-throughput analysis. As a proof of concept, we monitored the proliferation and differentiation of single Notch+ (stem-like) T47D breast cancer cells. The 3D collagen gel environment was found to be favorable for the stem-like phenotype through better self-renewal and de-differentiation (Notch− to Notch+ transition). More interestingly, we found that the Notch− de-differentiated cells were more resistant to doxorubicin and cisplatin than the Notch+ cells. Combining the 3D ECM culture and single cell resolution, the presented platform can automatically analyze the individual cell behaviors of hundreds of cells using a small amount of drug and reagents.