Engineered three-dimensional microfluidic device for interrogating cell-cell interactions in the tumor microenvironment

Abstract

Stromal cells in the tumor microenvironment play a key role in the metastatic properties of a tumor. It is recognized that cancer-associated fibroblasts (CAFs) and endothelial cells secrete factors capable of influencing tumor cell migration into the blood or lymphatic vessels. We developed a microfluidic device that can be used to image the interactions between stromal cells and tumor cell spheroids in a three dimensional (3D) microenvironment while enabling external control of interstitial flow at an interface, which supports endothelial cells. The apparatus couples a 200-lm channel with a semicircular well to mimic the interface of a blood vessel with the stroma, and the design allows for visualization of the interactions of interstitial flow, endothelial cells, leukocytes, and fibroblasts with the tumor cells. We observed that normal tissue-associated fibroblasts (NAFs) contribute to the "single file" pattern of migration of tumor cells from the spheroid in the 3D microenvironment. In contrast, CAFs induce a rapid dispersion of tumor cells out of the spheroid with migration into the 3D matrix. Moreover, treatment of tumor spheroid cultures with the chemokine CXCL12 mimics the effect of the CAFs, resulting in similar patterns of dispersal of the tumor cells from the spheroid. Conversely, addition of CXCL12 to co-cultures of NAFs with tumor spheroids did not mimic the effects observed with CAF co-cultures, suggesting that NAFs produce factors that stabilize the tumor spheroids to reduce their migration in response to CXCL12.

Microfluidic 3D cell culture: from tools to tissue models

“…Many parts within the metastatic cascade can be studied by co-culturing perfusable endothelial vessels with cancer cells. Using microfluidic 3D cell culture, effects such as angiogenesis [20 ,22 ,33], migration [49,50], intravasation [51] and extravasation [18,19,44,52] have been studied, and will be discussed in more detail. Figure 3c illustrates an extravasation setup in which endothelial cells are seeded against a compartmentalized hydrogel.…”

Study of Chemotaxis and Cell–Cell Interactions in Cancer with Microfluidic Devices

“…The chip was then etched in 10 M KOH at 80 °C for 1 h to form channels within the silica. Fabrication and assembly were conducted in an ISO 1000 stage clean room (Hockemeyer et al, 2014). Once the device is built, it can be reused several times for repeat experiments.…”

“…(G) 20 × magnification of DAPI channel visually confirms HMVECs are beginning to form a monolayer. Originally published in Hockemeyer et al (2014).…”

“…6) (Hockemeyer et al, 2014). This device provides an ideal tool to study these cell–cell interactions in a simulated tumor microenvironment.…”

Probing the Bi-directional Interaction Between Microglia and Gliomas in a Tumor Microenvironment on a Microdevice

“…Therefore, modeling the tumor microenvironment by integration of the interactions among different cell types on a micropatterned surface has become an attractive target of microfluidics. To this end, we and other groups have developed various microfluidic designs to study tumor invasion [16,17], tumorendothelial cell interactions [18], and tumor-stromal cell interactions [19].…”