We describe here designs of compartmentalized hydrogel microparticles with a tunable extracellular matrix (ECM) support for cell encapsulation and scalable 3D cell culture. The microparticles, rapidly formed by a one-step, multi-fluidic electrostatic spraying technique (>10 000 min À1 ), have a uniform spherical shape, a nearly monodisperse size distribution and controlled compartmentalization. They not only have a high surface area for mass transfer but also offer defined space and essential ECM support for various scalable and efficient 3D cell culture, co-culture and microtissue production applications.

Chemokine-mediated directed tumor cell migration within a three dimensional (3D) matrix, or chemoinvasion, is an important early step in cancer metastasis. Despite its clinical importance, it is largely unknown how cytokine and growth factor gradients within the tumor microenvironment regulate chemoinvasion. We studied tumor cell chemoinvasion in well-defined and stable chemical gradients using a robust 3D microfluidic model. We used CXCL12 (also known as SDF-1α) and epidermal growth factor (EGF), two well-known extracellular signaling molecules that co-exist in the tumor microenvironment (e.g. lymph nodes or intravasation sites), and a malignant breast tumor cell line, MDA-MB-231, embedded in type I collagen. When subjected to SDF-1α gradients alone, MDA-MB-231 cells migrated up the gradient, and the measured chemosensitivity (defined as the average cell velocity along the direction of the gradient) followed the ligand – receptor (SDF-1α – CXCR4) binding kinetics. On the other hand, when subjected to EGF gradients alone, tumor cells increased their overall motility, but without statistically significant chemotactic (directed) migration, in contrast to previous reports using 2D chemotaxis assays. Interestingly, we found that the chemoinvasive behavior to SDF-1α gradients was abrogated or even reversed in the presence of uniform concentrations of EGF; however, the presence of SDF-1α and EGF together modulated tumor cell motility cooperatively. These findings demonstrate the capabilities of our microfluidic model in re-creating complex microenvironments for cells, and the importance of cooperative roles of multiple cytokine and growth factor gradients in regulating cell migration in 3D environments.

The emerging field of micro-technology has opened up new possibilities for exploring cellular chemotaxis in real space and time, and at single cell resolution. Chemotactic cells sense and move in response to chemical gradients and play important roles in a number of physiological and pathological processes, including development, immune responses, and tumor cell invasions. Due to the size proximity of the microfluidic device to cells, microfluidic chemotaxis devices advance the traditional macro-scale chemotaxis assays in two major directions: one is to build well defined and stable chemical gradients at cellular length scales, and the other is to provide a platform for quantifying cellular responses at both cellular and molecular levels using advanced optical imaging systems. Here, we present a critical review on the designing principles, recent development, and potential capabilities of the microfluidic chemotaxis assay for solving problems that are of importance in the biomedical engineering field.