A customizable micropatterned platform for osteosarcoma spheroid generation, imaging, and drug screening

Area of Science:

• Biomedical Engineering
• Cancer Research
• Cell Biology

Background:

• Three-dimensional (3D) spheroids are valuable in vitro models for cancer drug screening, replicating tumor microenvironment and chemoresistance.
• Current spheroid culture methods struggle with automated real-time imaging and non-disruptive live image capture for treatment assessment.

Purpose of the Study:

• To develop an innovative platform for enhanced spheroid culture, enabling automated imaging and real-time analysis of drug responses.
• To overcome limitations in current spheroid culture systems for high-throughput drug screening and cancer research.

Main Methods:

• Fabrication of a custom culture dish with a micro-patterned agarose structure using 3D-printed molds.
• Immobilization of spheroids for automated high-throughput imaging with close proximity microscopy (micrometers).
• Compatibility testing with standard, confocal, and dual-photon microscopy, alongside viability and metabolic assays.

Main Results:

• Successful culture and treatment of osteosarcoma spheroids with chemotherapeutic agents.
• High-resolution confocal imaging of live spheroid extracellular matrix antigens.
• Demonstrated cost-effective, simultaneous imaging-based viability and metabolic assays with minimal reagent use.

Conclusions:

• The developed platform facilitates spheroid growth and immobilization, enabling automated data collection and analysis.
• This innovative tool supports in-depth investigation of spheroid behavior, therapeutic responses, and drug development.
• The platform allows tracking of spheroid size, shape, antigen expression, and viability over time.