Back to Organoid Biology: From Concept to Practice

Organoid Biology Course Modules

Module 1 — From Tissue Biopsy to Frozen Organoids

Overview:
This module provides a complete, practical introduction to generating organoids directly from patient or animal tissue.

Participants will learn:

  • Biopsy handling & processing: safe transfer, mechanical/enzymatic dissociation, viability assessment.
  • 3D matrix embedding: preparation and plating in Matrigel®
  • Medium formulation: selection and preparation of lineage-specific and disease-specific media.
  • Organoid maintenance: cleaning cultures, monitoring growth, feeding schedules, and troubleshooting.
  • Passaging: mechanical vs. enzymatic splitting and strategies for stable long-term expansion.
  • Culture validation: morphological checkpoints, viability assays, lineage markers, contamination control.
  • Cryopreservation: optimized freezing protocols, long-term storage, and post-thaw recovery principles.

Module 2 — From a Frozen Vial to a Growing Patient-Derived Organoid Culture

Overview:
This module teaches how to revive and expand organoids from a cryopreserved stock and prepare them for experimentation.

Participants will learn:

  • Correct thawing procedures and rapid recovery techniques.
  • Embedding thawed cells in ECM matrix and establishing early-stage cultures.
  • Medium selection, feeding schedules, and monitoring culture health.
  • Identifying and resolving early-growth challenges.

Module 3 — Forskolin-Induced Swelling (FIS) Assay: Functional Testing in Organoids

Overview:
Participants will learn to perform the Forskolin-Induced Swelling assay, one of the first functional assays developed for patient-derived intestinal organoids in cystic fibrosis (CF).

Participants will learn:

  • The biological principle: forskolin activation of CFTR → ion/water influx → organoid swelling.
  • Full experimental workflow: organoid preparation, forskolin exposure, imaging, and quantitative analysis.
  • Testing CFTR-modulating drugs and comparing responses between patient samples.
  • How swelling kinetics serve as a direct functional readout of CFTR activity and drug efficacy.

Module 4 — Drug Response in Cancer Organoids: Chemotherapy & Immunotherapy

Overview:
Participants will work with the cancer organoids they generated earlier in the course to perform drug-response experiments.

Participants will:

  • Design chemotherapy or immunotherapy assays, including optional co-culture with patient-matched or allogeneic immune cells.
  • Monitor organoid behavior in real time using advanced live-cell imaging (e.g., Incucyte).
  • Choose and perform appropriate analytical readouts based on their scientific question, such as:
    • flow cytometry
    • PCR or qPCR
    • mutation profiling
    • viability assays (XTT, CellTiter)
    • morphology-based analysis

Possible Additional Modules:

Module 5 — AI & Data-Driven Analysis in Organoid Biology

Overview: In this module, participants will learn how to transform raw organoid data—from images to multi-omics readouts—into quantitative insights that support treatment prediction and mechanistic interpretation. Rather than focusing on theory, this module provides practical, accessible tools for integrating microscopy, molecular assays, and clinical metadata using modern AI-assisted and data-analysis workflows.

Module 6 — Ethical & Regulatory Considerations for Human-Derived Materials in Organoid Research

Overview: In this module, participants will gain a clear understanding of the ethical, legal, and regulatory frameworks that govern the collection, handling, experimentation, and sharing of human-derived biological materials used for organoid culture. The session provides essential context for responsible research practice and ensures compliance with national and international standards.

  • Transfer Agreements (MTAs), and guidelines governing transport and international sharing of human samples and organoids.
  • Ethical challenges specific to organoids: issues related to long-term culture, genetic manipulation, creation of advanced models (e.g., brain organoids), and responsible communication of findings.