Droplet Array-Based 3D Coculture System for High-Throughput Tumor Angiogenesis Assay

Xiaohui Du, Wanming Li, Guansheng Du, Hansang Cho, Min Yu, Qun Fang, Luke P. Lee, Jin Fang

Research output: Contribution to journalArticlepeer-review

35 Scopus citations


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 × 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.

Original languageEnglish
Pages (from-to)3253-3261
Number of pages9
JournalAnalytical Chemistry
Issue number5
StatePublished - 6 Mar 2018

Bibliographical note

Funding Information:
We thank Jiabin Li for assistance with video production. This work was supported by National Natural Science Foundation of China (Grant nos. 81672920, 21375149, and 21435004).

Publisher Copyright:
© 2018 American Chemical Society.


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