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

Abstract

Microfluidic devices have very broad applications in biological assays from simple chemotaxis assays to much more complicated 3D bioreactors. In this chapter, we describe the design and methods for performing chemotaxis assays using simple microfluidic chemotaxis chambers. With these devices, using real-time video microscopy we can examine the chemotactic responses of neutrophil-like cells under conditions of varying gradient steepness or flow rate and then utilize software programs to calculate the speed and angles of cell migration as gradient steepness and flow are varied. Considering the shearing force generated on the cells by the constant flow that is required to produce and maintain a stable gradient, the trajectories of the cell migration will reflect the net result of both shear force generated by flow and the chemotactic force resulting from the chemokine gradient. Moreover, the effects of mutations in chemokine receptors or the presence of inhibitors of intracellular signals required for gradient sensing can be evaluated in real time. We also describe a method to monitor intracellular signals required for cells to alter cell polarity in response to an abrupt switch in gradient direction. Lastly, we demonstrate an in vitro method for studying the interactions of human cancer cells with human endothelial cells, fibroblasts, and leukocytes, as well as environmental chemokines and cytokines, using 3D microbioreactors that mimic the in vivo microenvironment.

A 3D Printed Device for Easy and Reliable Quantification of Fungal Chemotropic Growth

“…Unicellular organisms can be investigated quite easily with these systems: Liquid chemical gradients are established and can be changed by the utilization of pumps applying solutions from source to sink. The position of single cells can be tracked by fluorescence labeling, allowing retracing the movement reaction in detail (Wheeler et al, 2003;Choi et al, 2012;Sai et al, 2016). This is especially interesting for the investigation of tumor migration, since here even the invasion of tissue by single tumor cells can be monitored by applying different fluorescence markers for each cell type (Sai et al, 2016).…”

“…The position of single cells can be tracked by fluorescence labeling, allowing retracing the movement reaction in detail (Wheeler et al, 2003;Choi et al, 2012;Sai et al, 2016). This is especially interesting for the investigation of tumor migration, since here even the invasion of tissue by single tumor cells can be monitored by applying different fluorescence markers for each cell type (Sai et al, 2016). In yeast, microfluidic devices are used to monitor growth responses of mating projections (Brett et al, 2012).…”

Microfluidic and Paper-Based Devices for Disease Detection and Diagnostic Research

“…A microfluidic device developed by Sai et al facilitated the interrogation of cancer cells exposed to therapeutic drugs in the presence of other cell lines including stromal cells, immune cells, fibroblasts, and endothelial cells. They observed MDA-MB-342 cells sprouting into the surrounding ECM in the presence of CAFs [ 104 ]. Rogers et al reported on the interaction of MDA-MB-231 cells with normal-tissue associated fibroblasts in a 3D co-culture microfluidic system made to induce cellular migration.…”

Modeling alveolar injury using microfluidic co-cultures for monitoring bleomycin-induced epithelial/fibroblastic cross-talk disorder

“…Presently, cell migration assay based on 2D substrate has been allowed by various microuidic devices. 23,24 These devices can precisely congure chemoattractant gradients and allow quantitative cell analysis. 7,8 However, these approaches are not applicable to the migration analysis of interstitial cells (e.g.…”