Precise multiphase hydrogel engineering of miniaturized 3D cancer architectures via computationally informed microfluidics

Abstract

Understanding cancer biology and therapy responses requires accurate in vitro models that reflect tumor complexity. This work presents a multiphase microfluidic biofabrication approach for creating self-standing three-dimensional (3D) tumor models within hydrogel microfiber boundaries. A single framework enabled the fast generation of different in vitro cellular configurations, including discrete spheroids in size-limited liquid pockets and continuous multicellular fiberoids. These constructs incorporate key tumor features, including solid stress and microenvironmental interactions, which contribute to a more physiologically relevant replication of tumor responses. Computational simulations were used to fine-tune the biofabrication process, predicting fiber shapes and reducing costs associated with experimental iterations. In vitro tests demonstrated drug responsiveness in all configurations, with greatly enhanced manipulation of soft 3D cell structures. The fiberoid models further emulated intercellular dynamics herein explored in the glioblastoma-astrocyte context, expanding the versatility of our technology for cancer research, as a promising tool for drug discovery and precision-medicine strategies.