We report a noncytotoxic
resin compatible with and designed for
use in custom high-resolution 3D printers that follow the design approach
described in Gong et al., Lab Chip 17, 2899 (2017). The noncytotoxic
resin is based on a poly(ethylene glycol) diacrylate (PEGDA) monomer
with avobenzone as the UV absorber instead of 2-nitrophenyl phenyl
sulfide (NPS). Both NPS-PEGDA and avobenzone-PEGDA (A-PEGDA) resins
were evaluated for cytotoxicity and cell adhesion. We show that NPS-PEGDA
can be made effectively noncytotoxic with a postprint 12 h ethanol
wash, and that A-PEGDA, as-printed, is effectively noncytotoxic. 3D
prints made with either resin do not support strong cell adhesion
in their as-printed state; however, cell adhesion increases dramatically
with a short plasma treatment. Using A-PEGDA, we demonstrate spheroid
formation in ultralow adhesion 3D printed wells, and cell migration
from spheroids on plasma-treated adherent surfaces. Given that A-PEGDA
can be 3D printed with high resolution, it has significant promise
for a wide variety of cell-based applications using 3D printed microfluidic
structures.
The extracellular matrix (ECM) has pleiotropic effects, ranging from cell adhesion to cell survival. In tissue engineering, the use of ECM and ECM-like scaffolds has separated the field into two distinct areas—scaffold-based and scaffold-free. Scaffold-free techniques are used in creating reproducible cell aggregates which have massive potential for high-throughput, reproducible drug screening and disease modeling. Though, the lack of ECM prevents certain cells from surviving and proliferating. Thus, tissue engineers use scaffolds to mimic the native ECM and produce organotypic models which show more reliability in disease modeling. However, scaffold-based techniques come at a trade-off of reproducibility and throughput. To bridge the tissue engineering dichotomy, we posit that finding novel ways to incorporate the ECM in scaffold-free cultures can synergize these two disparate techniques.
Research in fields studying cellular response to surface tension and mechanical forces necessitate cell culture tools with tunability of substrate stiffness. We created a scalable hydrogel dish design to facilitate scaffold-free formation of multiple spheroids in a single dish. Our novel design features inner and outer walls, allowing efficient media changes and downstream experiments. The design is easily scalable, accommodating varying numbers of microwells per plate. We report that non-adherent hydrogel stiffness affects spheroid morphology and compaction. We found that spheroid morphology and viability in our hydrogel dishes were comparable to commercially available Aggrewell™800 plates, with improved tunability of surface stiffness and imaging area. Device function was demonstrated with a migration assay using two investigational inhibitors against EMT. We successfully maintained primary-derived spheroids from murine and porcine lungs in the hydrogel dish. These features increase the ability to produce highly consistent cell aggregates for biological research.
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