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Reproducible Manufacturing of SPOT as a High-throughput Scaffold-based Culture Platform.

Ruonan Cao1, Nancy T Li2, Chantelle B Shing1

  • 1Institute of Biomedical Engineering, University of Toronto.

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Patient-derived organoids (PDOs) offer a better way to model tumors. The Scaffold-supported Platform for Organoid-based Tissues (SPOT) device simplifies imaging and analysis for high-throughput drug screening in cancer research.

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Area of Science:

  • Biotechnology
  • Cancer Research
  • 3D Cell Culture

Background:

  • Patient-derived organoids (PDOs) are valuable cancer models due to their recapitulation of tumor complexity and heterogeneity.
  • Existing methods for organoid analysis can be cumbersome, limiting high-throughput applications.
  • There is a need for user-friendly devices to facilitate the analysis of organoid behavior.

Purpose of the Study:

  • To introduce the Scaffold-supported Platform for Organoid-based Tissues (SPOT) device for high-throughput 3D organoid culture.
  • To provide a detailed protocol for SPOT assembly, seeding, and quality control.
  • To enable streamlined imaging and analysis of organoids for cancer research.

Main Methods:

  • The SPOT platform is a 3D organoid culture device designed for high-throughput analysis.
  • It eliminates the meniscus formed by hydrogels, allowing for rapid, accurate imaging comparable to 2D cultures.
  • The protocol details fabrication in 96- and 384-well formats, compatible with manual and automated workflows.

Main Results:

  • SPOT facilitates easy, high-throughput imaging of organoids and co-cultures using standard microscopy.
  • The platform is accessible to researchers with basic tissue culture experience.
  • Fabrication is scalable, with multiple plates producible simultaneously.

Conclusions:

  • SPOT streamlines patient-derived organoid culture and analysis, enhancing their utility in research.
  • This device is a robust tool for high-throughput drug screening and translational cancer research.
  • SPOT simplifies the process of analyzing organoid behavior, accelerating discovery.