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Updated: Oct 18, 2025
A Brain Tumor/Organotypic Slice Co-culture System for Studying Tumor Microenvironment and Targeted Drug Therapies
Published on: November 7, 2015
Neuroblastoma and DIPG Organoid Coculture System for Personalized Assessment of Novel Anticancer Immunotherapies
Waleed M Kholosy1, Marc Derieppe1, Femke van den Ham1
1Princess Máxima Center for Paediatric Oncology, Heidelberglaan 25, 3584 CS Utrecht, The Netherlands.
Abstract:
Cancer immunotherapy has transformed the landscape of adult cancer treatment and holds a great promise to treat paediatric malignancies. However, in vitro test coculture systems to evaluate the efficacy of immunotherapies on representative paediatric tumour models are lacking. Here, we describe a detailed procedure for the establishment of an ex vivo test coculture system of paediatric tumour organoids and immune cells that enables assessment of different immunotherapy approaches in paediatric tumour organoids. We provide a step-by-step protocol for an efficient generation of patient-derived diffuse intrinsic pontine glioma (DIPG) and neuroblastoma organoids stably expressing eGFP-ffLuc transgenes using defined serum-free medium. In contrast to the chromium-release assay, the new platform allows for visualization, monitoring and robust quantification of tumour organoid cell cytotoxicity using a non-radioactive assay in real-time. To evaluate the utility of this system for drug testing in the paediatric immuno-oncology field, we tested our in vitro assay using a clinically used immunotherapy strategy for children with high-risk neuroblastoma, dinutuximab (anti-GD2 monoclonal antibody), on GD2 proficient and deficient patient-derived neuroblastoma organoids. We demonstrated the feasibility and sensitivity of our ex vivo coculture system using human immune cells and paediatric tumour organoids as ex vivo tumour models. Our study provides a novel platform for personalized testing of potential anticancer immunotherapies for aggressive paediatric cancers such as neuroblastoma and DIPG.
Insights
A new ex vivo system allows real-time testing of cancer immunotherapies on paediatric tumour organoids. This platform enables personalized treatment strategies for aggressive childhood cancers like neuroblastoma and diffuse intrinsic pontine glioma (DIPG).
Area of Science:
- Paediatric oncology
- Cancer immunotherapy
- Organoid technology
Background:
- Cancer immunotherapy has revolutionized adult cancer treatment but lacks effective in vitro models for paediatric malignancies.
- Existing methods for evaluating immunotherapy efficacy in paediatric tumours are insufficient.
Purpose of the Study:
- To establish a novel ex vivo coculture system for testing immunotherapies on paediatric tumour organoids.
- To enable real-time, non-radioactive assessment of immunotherapy efficacy in patient-derived paediatric tumour models.
Main Methods:
- Generation of patient-derived diffuse intrinsic pontine glioma (DIPG) and neuroblastoma organoids using defined serum-free medium.
- Development of an ex vivo coculture system with organoids and human immune cells for immunotherapy assessment.
- Utilisation of a real-time, non-radioactive assay for quantifying tumour organoid cell cytotoxicity.
- Testing the system with dinutuximab (anti-GD2 monoclonal antibody) on neuroblastoma organoids.
Main Results:
- Successful establishment of eGFP-ffLuc expressing DIPG and neuroblastoma organoids.
- Demonstration of a sensitive, real-time platform for monitoring immunotherapy-induced cytotoxicity.
- Validation of the system's feasibility and sensitivity using dinutuximab in a paediatric neuroblastoma model.
Conclusions:
- The developed ex vivo coculture system provides a novel platform for personalized immunotherapy testing in paediatric cancers.
- This system addresses the need for effective in vitro models in paediatric immuno-oncology.
- It holds promise for advancing treatment strategies for aggressive childhood cancers like neuroblastoma and DIPG.
