Project Objectives

• Validate air/liquid interface technology as a robust method for creating lung cancer organoids (LCOs) from patient biopsies.
• Compare organoids generated with conventional immersion culture versus this new method for success rate, homogeneity, cellular composition, histology, and genetic profile.
• Determine how closely organoids produced by each method replicate the original tumour using RNA sequencing.
• Adapt the technology for 24- and 96-well formats to support scalable drug-screening applications.
• Evaluate responses to standard chemotherapy and targeted therapies to assess suitability for personalised medicine and preclinical research.

3Rs Impact

• Provides a practical alternative to mouse-based lung cancer models, replacing patient-derived xenografts and genetically engineered or carcinogen-induced tumour models.
• Substantially reduces the need for animal use in drug-screening pipelines by enabling testing of up to 96 compounds per plate in vitro.
• Produces more reliable organoids, reducing follow-up in vivo studies that would otherwise be required to confirm results.
• Ensures better preservation of tumour microenvironment features, which improves scientific validity and reduces repeated experiments.
• Potential nationwide impact: estimated reduction of ~13,000 animals per year in Switzerland and ~250,000 per year across Europe for lung-cancer-related research.

Background

Lung cancer remains one of the most commonly diagnosed cancers worldwide and the leading cause of cancer-related deaths. Understanding tumour behaviour and predicting treatment responses require models that closely resemble a patient’s tumour. While organoids—miniature, patient-derived tumour models—are considered one of the best alternatives to animal experiments, generating high-quality lung cancer organoids (LCOs) remains difficult. Conventional methods grow organoids fully submerged in culture medium, a condition that does not reflect the lung’s natural exposure to air. This often results in low success rates, heterogeneous organoids, loss of key tumour features, and limited feasibility for drug screening.

This project addresses these limitations by validating a patented technology (3D-AirLiwell) that grows organoids in air/liquid interface conditions. This environment better mimics lung physiology and supports improved oxygen exchange, enhanced structural integrity, and better preservation of tumour microenvironment components. By comparing organoids produced with this new approach to those derived through standard immersion culture, the project aims to demonstrate improvements in success rate, homogeneity, molecular fidelity, and drug-response accuracy. Scaling the method to multi-well formats also enables high-throughput drug testing, which can support personalised medicine while reducing reliance on animal research.