Image-Assisted Microvessel-on-a-Chip Platform for Studying Cancer Cell Transendothelial Migration Dynamics

Abstract

With the push to reduce in vivo approaches, the demand for microphysiological models that recapitulate the in vivo settings in vitro is dramatically increasing. Here, we present an extracellular matrix-integrated microfluidic chip with a rounded microvessel of ~100 µm in diameter. Our system displays favorable characteristics for broad user adaptation: simplified procedure for vessel creation, minimised use of reagents and cells, and the ability to couple live-cell imaging and image analysis to study dynamics of cell-microenvironment interactions in 3D. Using this platform, the dynamic process of single breast cancer cells (LM2-4175) exiting the vessel lumen into the surrounding extracellular matrix was tracked. Here, we show that the presence of endothelial lining significantly reduced the cancer exit events over the 15-hour imaging period: there were either no cancer cells exiting, or the fraction of spontaneous exits was positively correlated with the number of cancer cells in proximity to the endothelial barrier. The capability to map the z-position of individual cancer cells within a 3D vessel lumen enabled us to observe cancer cell transmigration ‘hot spot’ dynamically. We also suggest the variations in the microvessel qualities may lead to the two distinct types of cancer transmigration behaviour. Our findings provide a tractable in vitro model applicable to other areas of microvascular research.

A 3D Printed Hanging Drop Dripper for Tumor Spheroids Analysis Without Recovery

“…To address this, researchers must recapitulate the tumor-endothelial model in the 3D scenario in vitro . Several microfluidic-based approaches have been developed to analyze tumor-endothelial cell interactions 44–46 . However, these methods are based on sophisticated microfabrication and require careful manipulation to construct the 3D interfaces between tumor and endothelial cells.…”

On-chip perivascularnichesupporting stemness of patient-derived glioma cells in a serum-free, flowable culture

“…The microfluidic device used to construct the microvessel-on-a-chip experiments was adapted from our previous design 22 and is schematically shown in Fig. 1 .…”

“…Shear stresses between 1.4 and 0.4 dyne per cm 2 were calculated within the side channel of the device with velocities around 1–2 mm s −1 as discussed previously. 22 …”

On-chipperivascular niche with patient-derived glioma cells

“…With the above goal, a simple microvessel-on-chip platform reported in our previous work 19 was adapted to support glioma-vessel formation. We describe the model design, the biological validation, and the use of the model as a microfluidic platform as a tool to study the cellular behaviour and interactions indeed differ from those tracked within non-tissue specific vascular niches (i.e.…”