3D Printed Solutions for Spheroid Engineering and Cancer Research

Area of Science:

• Biotechnology
• 3D Bioprinting
• Cancer Biology

Background:

• Multicellular organisms rely on 3D cellular organization for organogenesis and tissue development.
• 3D cell culture models offer physiologically relevant insights into development and cancer biology, unattainable in 2D cultures.
• Cellular spheroids are crucial in vitro tumor models, typically generated using the hanging drop method.

Purpose of the Study:

• To present a novel 3D printing approach for fabricating custom hanging drop devices for tumor cell culture.
• To design and create modularized, high-throughput hanging drop devices (Q-serts) for efficient spheroid formation and automation.
• To establish the utility of these devices in generating synthetic tumor microenvironment mimics (STEMs) using human cancer cells.

Main Methods:

• Fabrication of custom hanging drop devices (Q-serts) using fused filament deposition (FFD).
• Engineering of unicellular and multicellular spheroids (STEMs) using human cancer cells.
• Automation of spheroid culture using a pipetting robot and bioprinting with a custom carboxylated agarose bioink to simulate the tumor microenvironment (TME).
• Characterization of spheroids via light microscopy and histology.

Main Results:

• Demonstrated successful fabrication of bespoke, modularized hanging drop devices (Q-serts) via 3D printing.
• Established the efficacy of Q-serts in forming both unicellular and multicellular spheroids (STEMs) with high morphological and structural integrity.
• Confirmed high cell viability throughout the automated spheroid culture and bioprinting workflow.
• Validated the use of a custom bioink for simulating the tumor microenvironment (TME).

Conclusions:

• The presented 3D printing-driven platform offers a user-focused, reproducible, and scalable solution for spheroid culture.
• Automated spheroid preparation and culture using these devices minimize user-dependent variables, improving standardization.
• This technology has the potential to advance translational cancer research by enabling direct comparison of findings.
• The developed system provides a robust method for creating physiologically relevant tumor microenvironment mimics (STEMs).