A 3D Bioprinted in vitro Model of Neuroblastoma Recapitulates Dynamic Tumor‐Endothelial Cell Interactions Contributing to Solid Tumor Aggressive Behavior

Abstract

Neuroblastoma (NB) is the most common extracranial tumor in children resulting in substantial morbidity and mortality. A deeper understanding of the NB tumor microenvironment (TME) remains an area of active research but there is a lack of reliable and biomimetic experimental models. This study utilizes a 3D bioprinting approach, in combination with NB spheroids, to create an in vitro vascular model of NB for exploring the tumor function within an endothelialized microenvironment. A gelatin methacryloyl (gelMA) bioink is used to create multi‐channel cubic tumor analogues with high printing fidelity and mechanical tunability. Human‐derived NB spheroids and human umbilical vein endothelial cells (HUVECs) are incorporated into the biomanufactured gelMA and cocultured under static versus dynamic conditions, demonstrating high levels of survival and growth. Quantification of NB‐EC integration and tumor cell migration suggested an increased aggressive behavior of NB when cultured in bioprinted endothelialized models, when cocultured with HUVECs, and also as a result of dynamic culture. This model also allowed for the assessment of metabolic, cytokine, and gene expression profiles of NB spheroids under varying TME conditions. These results establish a high throughput research enabling platform to study the TME‐mediated cellular‐molecular mechanisms of tumor growth, aggression, and response to therapy.

3Din vitromodeling of the exocrine pancreatic unit using tomographic volumetric bioprinting

“…3a,b). This is in line with other works, which have cultured viable tomographically printed constructs for several weeks 46,51,66 . Therefore, the results proved the beneficial effects given by GelMA as bioink, matching with previous reports on the extensive use of this material in biomedical applications 67–71 .…”

3D bioprinting of nanoparticle-laden hydrogel scaffolds with enhanced antibacterial and imaging properties

“…A 0.5% (w/v) concentration of lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) was used as a photoinitiator to crosslink the bioinks under a 10 mW/cm 2 UV light source for 2 min. These crosslinking parameters were chosen based on our previous experiences ( Li et al., 2022 ; ML Tomov et al., 2019 ; Ning et al., 2020 ; Ning et al., 2022 ; Ning et al., 2021 ) to generate GelMA constructs at biologically relevant stiffness (mimicking soft tissue stiffness, ∼20–140 kPa ( Huang et al., 2012 ; Serpooshan et al., 2013 ; Wei et al., 2015 )). 3D printed constructs were designed using Autodesk Fusion 360 CAD software (Autodesk Inc., San Rafael, CA) and converted to the standard STL file format for printing.…”

Advances in Microfluidics: Technical Innovations and Applications in Diagnostics and Therapeutics

“…Lymphatic muscle cells aligned circumferentially while endothelial cells aligned axially under flow, as only observed in vivo in the past. In another instance, a 3D bioprinting approach, in combination with neuroblastoma (NB) spheroids, generated NB vascularization, allowing one to explore the tumor function within an endothelialized microenvironment …”

“…In another instance, a 3D bioprinting approach, in combination with neuroblastoma (NB) spheroids, generated NB vascularization, allowing one to explore the tumor function within an endothelialized microenvironment. 137 New fields beyond neuroscience, immunology, or cardiology may benefit from tissue-and organ-on-chip technology. Obesity and associated diseases, such as diabetes, are recent examples.…”