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.

The Role of Microenvironmental Cues and Mechanical Loading Milieus in Breast Cancer Cell Progression and Metastasis

“…Taking the perspective of tumors as interconnected organ systems, further work is clearly required to develop model tumor systems that provide realistic cell-cell interactions and recapitulate the response of multiple cell types to mechanical signals (Bregenzer et al, 2019). For example, one study recapitulated 3D blood vessels with a parallel epithelial-lined lumen to investigate the epithelial-endothelial migration in breast cancer (Devadas et al, 2019). This is of significance since the endothelial-epithelial interactions are not well-understood yet for 3D environments.…”

“…Microfluidic platforms can provide an easily customizable solution for simulating tumor environments, for example, the microfluidic parallel lumen device targeting endothelial and epithelial co-culture as described above (Devadas et al, 2019). Devices may further be customized for each segment of the metastatic process.…”

Reversible Bonding of Thermoplastic Elastomers for Cell Patterning Applications

“…Recently, microfluidic-based models have emerged as powerful platforms to study cell migration and/or angiogenesis [5]. The chipbased models enable formation of 3D architecture blood vessels and the study of soluble biochemistry in discrete fluidic channel networks [6], as well as the study of epithelialendothelial migration in breast cancer [7]. These novel microfluidic approaches closely mimic both physiological and pathological microenvironments and there is a significant effort to apply these technologies to new drug screening and therapeutic applications.…”

“…These novel microfluidic approaches closely mimic both physiological and pathological microenvironments and there is a significant effort to apply these technologies to new drug screening and therapeutic applications. However, microfluidic device fabrication usually involves complicated cleanroom facilities and requires highly sophisticated processes which are costly and labor-intensive [6][7][8][9]. Moreover, poly(dimethylsiloxane) (PDMS) is a commonly used material in the prototype of microfluidic devices.…”

“…TP materials are well-known cell culture substrates. Due to their high biocompatibility, low-cytotoxicity, minimal drugs absorption and high optical transparency, they have been used extensively in 3D cell culture platforms [7,13], as biocompatible substrates for cellular contact guidance [14], lab-on-a-chip devices [15], and other biomedical applications generally [16]. A subset class of TP materials called thermoplastic elastomers (TPEs) are also soft and flexible offering distinct advantages in manufacturing processes.…”