Heterogeneous spheroids with tunable interior morphologies by droplet-based microfluidics

Heterogeneous spheroids that mimic the complex three-dimensional environment of natural tissues are needed in various biomedical applications. Geometric cues from cellular matrix play invaluable roles in governing cell behavior and phenotype. However, the structural complexity of interior morphologies of spheroids is currently limited due to poor spatial resolution of positioning/orientation of cellular constructs. Here, a coaxial capillary microfluidic device is developed to generate gelatin methacrylate (GelMA) microspheres with tunable dimensions and interior morphologies, such as core-shell, or microspheres with interior undulated wavy, or spiral canals, by manipulating the two-phase flow of hydrogel precursor solution and methylcellulose solution. The formation of diverse and exquisite interior morphologies is caused by the interacting viscous instabilities of the two-phase flow in the microfluidic system, followed by water-in-oil emulsion and photo-initiated polymerization. Polyethylene glycol diacrylate (PEGDA) is incorporated into the GelMA solution to tune the mechanical properties of the fabricated microspheres, and an optimized concentration of PEGDA is confirmed by evaluating the in vitro proliferation and vascularization of human umbilical vein endothelial cells. Further, a heterogeneous spheroid with spiral blood vessel lumen is constructed to demonstrate the versatility and potential of the proposed droplet-based microfluidic approach for building functional tissue constructs.