Abstract
Hypoxia plays a central role in cancer progression and resistance to therapy. We have engineered a microdevice platform to recapitulate the intratumor oxygen gradients that drive the heterogeneous hypoxic landscapes in solid tumors. Our design features a “tumor section”-like culture by incorporating a cell layer between two diffusion barriers, where an oxygen gradient is established by cellular metabolism and physical constraints. We confirmed the oxygen gradient by numerical simulation and imaging-based oxygen sensor measurement. We also demonstrated spatially-resolved hypoxic signaling in cancer cells through immunostaining, gene expression assay, and hypoxia-targeted drug treatment. Our platform can accurately generate and control oxygen gradients, eliminates complex microfluidic handling, allows for incorporation of additional tumor components, and is compatible with high-content imaging and high-throughput applications. It is well suited for understanding hypoxia-mediated mechanisms in cancer disease and other biological processes, and discovery of new therapeutics.
Highlights
Considerable efforts have been made to establish hypoxic tumor models that can be analyzed with ease and reproducibility
When we analyze the differential gene expression between pillar center (PC) and pillar edge (PE) in the same samples, we discovered that the gene expression in the cells from the hypoxia device are spatially regulated by the oxygen gradient, with down-regulated proliferation and up-regulated markers in PC for apoptosis, glycolysis and migration/metastasis
The lateral oxygen gradient created on a monolayer cell culture allows for real-time, high-content investigation of cellular phenotypes and behaviors with wide-field microscopy-based techniques, as demonstrated by the laser capture microdissection (LCM)-based gene expression analysis
Summary
Considerable efforts have been made to establish hypoxic tumor models that can be analyzed with ease and reproducibility. Available hypoxia chambers provide one oxygen concentration at a time, limiting its throughput in testing cell responses to different oxygen levels These approaches fail to capture the spatial complexity of oxygen profiles and the resulted crosstalk in a hypoxic tumor[22,23]. Paper-supported 3D cell cultures have been developed to recapitulate gradients in spheroids and tumors, where layers of 2D cultures are stacked to establish the gradients, and disassembled for imaging and analysis[27] Such methods lack a lateral gradient profile for microscopy, and require additional handling to analyze cells on each layer. We present the device and platform as a versatile tool for gaining insights into cancer biology and accelerate the development and discovery of new therapeutics
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