A microfluidic mammary gland coculture model using parallel 3D lumens for studying epithelial-endothelial migration in breast cancer

Abstract

In breast cancer development, crosstalk between mammary epithelial cells and neighboring vascular endothelial cells is critical to understanding tumor progression and metastasis, but the mechanisms of this dynamic interplay are not fully understood. Current cell culture platforms do not accurately recapitulate the 3D luminal architecture of mammary gland elements. Here, we present the development of an accessible and scalable microfluidic coculture system that incorporates two parallel 3D luminal structures that mimic vascular endothelial and mammary epithelial cell layers, respectively. This parallel 3D lumen configuration allows investigation of endothelial-epithelial crosstalk and its effects of the comigration of endothelial and epithelial cells into microscale migration ports located between the parallel lumens. We describe the development and application of our platform, demonstrate generation of 3D luminal cell layers for endothelial cells and three different breast cancer cell lines, and quantify their migration profiles based on number of migrated cells, area coverage by migrated cells, and distance traveled by individual migrating cells into the migration ports. Our system enables analysis at the single-cell level, allows simultaneous monitoring of endothelial and epithelial cell migration within a 3D extracellular matrix, and has potential for applications in basic research on cellular crosstalk as well as drug development.