Droplet Array-Based 3D Coculture System for High-Throughput Tumor Angiogenesis Assay
Droplet Array-Based 3D Coculture System for High-Throughput Tumor Angiogenesis Assay
复制标题
基于液滴阵列的 3D 共培养系统，用于高通量肿瘤血管生成检测
DOI：
10.1021/acs.analchem.7b04772
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发表时间：
2018-03-06
影响因子：
7.4
通讯作者：
Fang, Jin
中科院分区：
文献类型：
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作者：
Du, Xiaohui;Li, Wanming;Fang, Jin
血管生成是肿瘤进展和转移的关键，它通过多细胞相互作用进行。因此，迫切需要高通量的肿瘤血管生成检测，同时检查多个因素。为了研究肿瘤血管生成，我们开发了一种基于微流控液滴阵列的细胞共培养系统，该系统包括具有6 × 9对孔阵列的双层聚二甲基硅氧烷芯片和自动化液滴操作装置。在每个液滴对单元中，通过将细胞悬浮液与基质胶混合，在一个液滴中以3D培养肿瘤细胞，在另一个液滴中，以2D培养人脐静脉内皮细胞（HUVEC）。通过一种新开发的融合方法融合液滴，并通过在融合的液滴单元中共培养肿瘤细胞和HUVECs来检测肿瘤血管生成。3D培养的肿瘤细胞形成聚集体，体内观察到缺氧中心，并分泌更多的血管内皮生长因子（VEGF）和更强烈地诱导HUVEC小管形成比2D培养的肿瘤细胞。我们的单个阵列支持54个平行测定。不同肿瘤细胞的血管生成潜力及其对抗血管生成剂Fingolimod的不同反应可以在单个阵列中无相互干扰地进行研究。我们的液滴为基础的测定是方便的，以评估多细胞相互作用在高通量的背景下，肿瘤萌芽血管生成，我们设想，该测定可以广泛实施研究其他细胞的细胞相互作用。
Angiogenesis is critical for tumor progression and metastasis, and it progresses through orchestral multicellular interactions. Thus, there is urgent demand for high-throughput tumor angiogenesis assays for concurrent examination of multiple factors. For investigating tumor angiogenesis, we developed a microfluidic droplet array-based cell-coculture system comprising a two-layer polydimethylsiloxane chip featuring 6 x 9 paired-well arrays and an automated droplet manipulation device. In each droplet-pair unit, tumor cells were cultured in 3D in one droplet by mixing cell suspensions with Matrigel, and in the other droplet, human umbilical vein endothelial cells (HUVECs) were cultured in 2D. Droplets were fused by a newly developed fusion method, and tumor angiogenesis was assayed by coculturing tumor cells and HUVECs in the fused droplet units. The 3D-cultured tumor cells formed aggregates harboring a hypoxic center as observed in vivo and secreted more vascular endothelial growth factor (VEGF) and more strongly induced HUVEC tubule formation than did 2D-cultured tumor cells. Our single array supported 54 assays in parallel. The angiogenic potentials of distinct tumor cells and their differential responses to antiangiogenesis agent, Fingolimod, could be investigated without mutual interference in a single array. Our droplet-based assay is convenient to evaluate multicellular interaction in high throughput in the context of tumor sprouting angiogenesis, and we envision that the assay can be extensively implementable for studying other cell cell interactions.