Biofabrication of Modular Spheroids as Tumor-Scale Microenvironments for Drug Screening

To streamline the drug discovery pipeline, there is a pressing need for preclinical models which replicate the complexity and scale of native tumors. While there have been advancements in the formation of microscale tumor units, these models are cell-line dependent, time-consuming and have not improved clinical trial success rates. In this study, two methods for generating 3D tumor microenvironments are compared, rapidly fabricated hydrogel microspheres and traditional cell-dense spheroids. These modules are then bioassembled into 3D printed thermoplastic scaffolds, using an automated biofabrication process, to form tumor-scale models. Modules are formed with SKOV3 and HFF cells as monocultures and cocultures, and the fabrication efficiency, cell architecture, and drug response profiles are characterized, both as single modules and as multimodular constructs. Cell-encapsulated Gel-MA microspheres are fabricated with high-reproducibility and dimensions necessary for automated tumor-scale bioassembly regardless of cell type, however, only cocultured spheroids form compact modules suitable for bioassembly. Chemosensitivity assays demonstrate the reduced potency of doxorubicin in coculture bioassembled constructs and a approximate to five-fold increase in drug resistance of cocultured cells in 3D modules compared with 2D monolayers. This bioassembly system is efficient and tailorable so that a variety of relevant-sized tumor constructs could be developed to study tumorigenesis and modernize drug discovery.

Automated summary

In order to improve the drug discovery process, there is a need for preclinical models that can mimic the complexity and scale of native tumors. This study compares two methods for creating 3D tumor microenvironments, hydrogel microspheres and cell-dense spheroids, which are then assembled into 3D printed scaffolds to form tumor-scale models. The efficiency, cell architecture, and drug response profiles of these models are characterized, and it is found that only cocultured spheroids are suitable for bioassembly. The bioassembly system demonstrates high efficiency and customization, making it valuable for studying tumorigenesis and advancing drug discovery.